Modulating plant sugar levels

ABSTRACT

This document provides methods and materials related to plants having modulated (e.g., increased) levels of sugars (e.g., glucose, fructose, and/or sucrose). For example, this document provides plants having increased sugar levels as well as methods and materials for making plants and plant products having increased sugar levels.

CROSS-REFERENCE TO RELATED APPLICATIONS

This Application claims priority under 35 U.S.C. 119 to U.S. ProvisionalApplication No. 60/610,356, filed Sep. 14, 2004, incorporated herein byreference in its entirety.

TECHNICAL FIELD

This document provides methods and materials related to plants havingmodulated (e.g., increased) levels of sugars (e.g., glucose, fructose,and/or sucrose). For example, this document provides plants havingincreased sugar levels as well as methods and materials for makingplants and plant products having increased sugar levels.

BACKGROUND

A sugar is a carbohydrate that is sweet to taste. Sugars are used infood and drink as a source of sweetness and energy and are important inbiochemistry. Sucrose, also called “table sugar,” is a white crystallinesolid. Sucrose is a disaccharide composed of two monosaccharides,glucose and fructose, joined together by a 1→2-α,β-glycosidic bond.Sucrose is commercially extracted from either sugar cane or sugar beetand then purified and crystallized. Other commercial sources aresorghum, date palm, and sugar maples. The monosaccharides, such asglucose (which is produced from sucrose by enzymes or acid hydrolysis),are a store of energy that is used by biological cells. Oxidation ofglucose is known as glycolysis. It occurs in virtually all cells.Glucose is oxidized to either lactate or pyruvate. Under aerobicconditions, the dominant product in most tissues is pyruvate and thepathway is known as aerobic glycolysis. When oxygen is depleted, as forinstance during prolonged vigorous exercise, the dominant glycolyticproduct in many tissues is lactate and the process is known as anaerobicglycolysis. Other sugars besides glucose, such as fructose, can enterglycolysis after being converted to appropriate intermediates that canenter the pathway. Glycolysis results in production of NADH and ATP. TheNADH generated during glycolysis is used to fuel mitochondrial ATPsynthesis via oxidative phosphorylation. ATP powers virtually everyactivity of the cell and organism. Organisms from the simplest bacteriato humans use ATP as their primary energy currency.

SUMMARY

This document provides methods and materials related to modulating sugarlevels in plants. For example, this document provides plants havingincreased levels of sugars, plant cells and seeds having the ability togrow into plants having increased levels of sugars, plant products(e.g., sugar extracts, sugar syrup, molasses, food, foodstuffs, andanimal feed) having increased levels of sugars, and methods for makingsuch plants, plant cells, and plant products. Plants having the abilityto produce increased levels of sugars can be used as sugar sources orsources of plant products having increased levels of sugars. Forexample, making sugars from plants having the ability to produceincreased levels of sugars can allow sugar manufacturers to increasesugar yields (e.g., tons of sugar per acre). In addition, plants andplant products having increased levels of sugars can be used as foods oringredients in food products having increased nutritional value andflavor per serving.

In one embodiment, a method of modulating the level of a sugar in aplant is provided. The method can include introducing into a plant cellan isolated nucleic acid including a nucleic acid sequence encoding apolypeptide having 80 percent or greater sequence identity to an aminoacid sequence selected from the group consisting of SEQ ID NO:14, SEQ IDNO:2, SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4,Ceres clone SEQ ID NO:9, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ IDNO:8, and the Consensus sequence set forth in FIG. 6, where a plantproduced from the plant cell has a different sugar level as compared toa sugar level in a corresponding control plant that does not include theisolated nucleic acid. The sequence identity can be 85 percent orgreater, 90 percent or greater, or 95 percent or greater.

In another embodiment, a method of modulating the level of a sugar in aplant is provided. The method can include introducing into a plant cellan isolated nucleic acid including a nucleic acid sequence encoding apolypeptide having 80 percent or greater sequence identity to an aminoacid sequence selected from the group consisting of SEQ ID NO:14, SEQ IDNO:2, SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4,SEQ ID NO:9, and the Consensus sequence set forth in FIG. 6, where aplant produced from the plant cell has a different sugar level ascompared to a sugar level in a corresponding control plant that does notinclude the isolated nucleic acid. The sequence identity can be 85percent or greater, 90 percent or greater, or 95 percent or greater.

In another embodiment, a method of modulating the level of a sugar in aplant is provided. The method can include introducing into a plant cellan isolated nucleic acid including a nucleic acid sequence encoding apolypeptide having 80 percent or greater sequence identity to an aminoacid sequence selected from the group consisting of SEQ ID NO:14, SEQ IDNO:2, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:4, and the Consensus sequenceset forth in FIG. 6, where a plant produced from the plant cell has adifferent sugar level as compared to a sugar level in a correspondingcontrol plant that does not include the isolated nucleic acid. Thesequence identity can be 85 percent or greater, 90 percent or greater,or 95 percent or greater.

In a further embodiment, a method of modulating the level of a sugar ina plant is provided. The method can include introducing into a plantcell an isolated nucleic acid including a nucleic acid sequence encodinga polypeptide having 80 percent or greater sequence identity to an aminoacid sequence selected from the group consisting of SEQ ID NO:14, SEQ IDNO:2, SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:4, where a plant producedfrom the plant cell has a different sugar level as compared to a sugarlevel in a corresponding control plant that does not include theisolated nucleic acid. The sequence identity can be 85 percent orgreater, 90 percent or greater, or 95 percent or greater.

In another aspect, a method of modulating the level of a sugar in aplant is provided. The method can include introducing into a plant cellan isolated nucleic acid including a nucleic acid sequence encoding apolypeptide including an amino acid sequence corresponding to SEQ IDNO:2, where a plant produced from the plant cell has a different sugarlevel as compared to a sugar level in a corresponding control plant thatdoes not include the isolated nucleic acid.

In another aspect, a method of modulating the level of a sugar in aplant is provided. The method can include introducing into a plant cellan isolated nucleic acid including a nucleic acid sequence encoding apolypeptide including an amino acid sequence corresponding to SEQ IDNO:14, where a plant produced from the plant cell has a different sugarlevel as compared to a sugar level in a corresponding control plant thatdoes not include the isolated nucleic acid.

In yet another aspect, a method of modulating the level of a sugar in aplant is provided. The method can include introducing into a plant cellan isolated nucleic acid including a nucleic acid sequence encoding apolypeptide including an amino acid sequence corresponding to theConsensus sequence set forth in FIG. 6, where a plant produced from theplant cell has a different sugar level as compared to a sugar level in acorresponding control plant that does not include the isolated nucleicacid.

In another embodiment, a method of modulating the level of a sugar in aplant is provided. The method can include introducing into a plant cell(a) a first isolated nucleic acid including a nucleic acid sequenceencoding a polypeptide having 80 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9,SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, andthe Consensus sequence set forth in FIG. 6; and (b) a second isolatednucleic acid including a nucleic acid sequence encoding a polypeptidehaving 80 percent or greater sequence identity to an amino acid sequencecorresponding to SEQ ID NO:14; where a plant produced from the plantcell has a different sugar level as compared to a sugar level in acorresponding control plant that does not include the first isolatednucleic acid or the second isolated nucleic acid.

In a further embodiment, a method of modulating the level of a sugar ina plant is provided. The method can include introducing into a plantcell (a) a first isolated nucleic acid including a nucleic acid sequenceencoding a polypeptide including an amino acid sequence selected fromthe group consisting of SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ IDNO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12,SEQ ID NO:3, SEQ ID NO:8, and the Consensus sequence set forth in FIG.6; and (b) a second isolated nucleic acid including a nucleic acidsequence encoding a polypeptide including an amino acid sequencecorresponding to SEQ ID NO:14; where a plant produced from the plantcell has a different sugar level as compared to a sugar level in acorresponding control plant that does not include the first isolatednucleic acid or the second isolated nucleic acid.

In another embodiment, a method of modulating the level of a sugar in aplant is provided. The method can include introducing into a plant cell(a) a first isolated nucleic acid including a nucleic acid sequenceencoding a polypeptide having 80 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9,SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, and SEQ ID NO:8;and (b) a second isolated nucleic acid including a nucleic acid sequenceencoding a polypeptide having 80 percent or greater sequence identity toan amino acid sequence corresponding to SEQ ID NO:14; where a plantproduced from the plant cell has a different sugar level as compared toa sugar level in a corresponding control plant that does not include thefirst isolated nucleic acid or the second isolated nucleic acid.

In yet another embodiment, a method of modulating the level of a sugarin a plant is provided. The method can include introducing into a plantcell (a) a first isolated nucleic acid including a nucleic acid sequenceencoding a polypeptide including an amino acid sequence selected fromthe group consisting of SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ IDNO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12,SEQ ID NO:3, and SEQ ID NO:8; and (b) a second isolated nucleic acidincluding a nucleic acid sequence encoding a polypeptide including anamino acid sequence corresponding to SEQ ID NO:14; where a plantproduced from the plant cell has a different sugar level as compared toa sugar level in a corresponding control plant that does not include thefirst isolated nucleic acid or the second isolated nucleic acid.

A different sugar level can be an increased level of one or more sugars,such as glucose, fructose, or sucrose. A different sugar level can be anincreased level of glucose and fructose, or an increased level ofglucose, fructose, and sucrose.

In another embodiment, a method of producing a plant having a modulatedlevel of a sugar is provided. The method can include (a) introducinginto a plant cell an isolated nucleic acid including a nucleic acidsequence encoding a polypeptide having 80 percent or greater sequenceidentity to an amino acid sequence selected from the group consisting ofSEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:7, SEQ ID NO:12, SEQ IDNO:3, SEQ ID NO:8, and the Consensus sequence set forth in FIG. 6; and(b) growing a plant from the plant cell. The sequence identity can be 85percent or greater, 90 percent or greater, or 95 percent or greater.

In another embodiment, a method of producing a plant having a modulatedlevel of a sugar is provided. The method can include (a) introducinginto a plant cell an isolated nucleic acid including a nucleic acidsequence encoding a polypeptide having 80 percent or greater sequenceidentity to an amino acid sequence selected from the group consisting ofSEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQID NO:11, SEQ ID NO:4, SEQ ID NO:9, and the Consensus sequence set forthin FIG. 6; and (b) growing a plant from the plant cell. The sequenceidentity can be 85 percent or greater, 90 percent or greater, or 95percent or greater.

In a further embodiment, a method of producing a plant having amodulated level of a sugar is provided. The method can include (a)introducing into a plant cell an isolated nucleic acid including anucleic acid sequence encoding a polypeptide having 80 percent orgreater sequence identity to an amino acid sequence selected from thegroup consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6,SEQ ID NO:4, and the Consensus sequence set forth in FIG. 6; and (b)growing a plant from the plant cell. The sequence identity can be 85percent or greater, 90 percent or greater, or 95 percent or greater.

In another aspect, a method of producing a plant having a modulatedlevel of a sugar is provided. The method can include (a) introducinginto a plant cell an isolated nucleic acid including a nucleic acidsequence encoding a polypeptide having 80 percent or greater sequenceidentity to an amino acid sequence selected from the group consisting ofSEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:4;and (b) growing a plant from the plant cell. The sequence identity canbe 85 percent or greater, 90 percent or greater, or 95 percent orgreater.

In another aspect, a method of producing a plant having a modulatedlevel of a sugar is provided. The method can include (a) introducinginto a plant cell an isolated nucleic acid including a nucleic acidsequence encoding a polypeptide including an amino acid sequencecorresponding to SEQ ID NO:14; and (b) growing a plant from the plantcell.

In another aspect, a method of producing a plant having a modulatedlevel of a sugar is provided. The method can include (a) introducinginto a plant cell an isolated nucleic acid including a nucleic acidsequence encoding a polypeptide including an amino acid sequencecorresponding to SEQ ID NO:2; and (b) growing a plant from the plantcell.

In yet another aspect, a method of producing a plant having a modulatedlevel of a sugar is provided. The method can include (a) introducinginto a plant cell an isolated nucleic acid including a nucleic acidsequence encoding a polypeptide including an amino acid sequencecorresponding to the Consensus sequence set forth in FIG. 6; and (b)growing a plant from the plant cell.

In another embodiment, a method of producing a plant having a modulatedlevel of a sugar is provided. The method can include (a) introducinginto a plant cell a first isolated nucleic acid including a nucleic acidsequence encoding a polypeptide having 80 percent or greater sequenceidentity to an amino acid sequence selected from the group consisting ofSEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ IDNO:8, and the Consensus sequence set forth in FIG. 6, and a secondisolated nucleic acid including a nucleic acid sequence encoding apolypeptide having 80 percent or greater sequence identity to an aminoacid sequence corresponding to SEQ ID NO:14; and (b) growing a plantfrom the plant cell.

In a further embodiment, a method of producing a plant having amodulated level of a sugar is provided. The method can include (a)introducing into a plant cell a first isolated nucleic acid including anucleic acid sequence encoding a polypeptide including an amino acidsequence selected from the group consisting of SEQ ID NO:5, SEQ IDNO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:2,SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, and the Consensussequence set forth in FIG. 6, and a second isolated nucleic acidincluding a nucleic acid sequence encoding a polypeptide including anamino acid sequence corresponding to SEQ ID NO:14; and (b) growing aplant from the plant cell.

In another embodiment, a method of producing a plant having a modulatedlevel of a sugar is provided. The method can include (a) introducinginto a plant cell a first isolated nucleic acid including a nucleic acidsequence encoding a polypeptide having 80 percent or greater sequenceidentity to an amino acid sequence selected from the group consisting ofSEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, and SEQ IDNO:8, and a second isolated nucleic acid including a nucleic acidsequence encoding a polypeptide having 80 percent or greater sequenceidentity to an amino acid sequence corresponding to SEQ ID NO:14; and(b) growing a plant from the plant cell.

In yet another embodiment, a method of producing a plant having amodulated level of a sugar is provided. The method can include (a)introducing into a plant cell a first isolated nucleic acid including anucleic acid sequence encoding a polypeptide including an amino acidsequence selected from the group consisting of SEQ ID NO:5, SEQ IDNO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:2,SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, and SEQ ID NO:8, and a secondisolated nucleic acid including a nucleic acid sequence encoding apolypeptide including an amino acid sequence corresponding to SEQ IDNO:14; and (b) growing a plant from the plant cell.

A modulated sugar level can be an increased level of one or more sugars,such as glucose, fructose, or sucrose. A modulated sugar level can be anincreased level of glucose and fructose, or an increased level ofglucose, fructose, and sucrose.

An isolated nucleic acid or an exogenous nucleic acid can be operablylinked to a regulatory region, such as a promoter. The promoter can be abroadly expressing promoter, such as p326, YP0158, YP0214, YP0380,PT0848, PT0633, YP0050, YP0144, or YP0190. A promoter can be acell-specific or tissue-specific promoter, such as a seed-specificpromoter, a root-specific promoter, or a non-seed fruit tissue promoter.A seed-specific promoter can be the napin promoter, the Arcelin-5promoter, the phaseolin gene promoter, the soybean trypsin inhibitorpromoter, the ACP promoter, the stearoyl-ACP desaturase gene, thesoybean α subunit of β-conglycinin promoter, the oleosin promoter, the15 kD zein promoter, the 16 kD zein promoter, the 19 kD zein promoter,the 22 kD zein promoter, the 27 kD zein promoter, the Osgt-1 promoter,the beta-amylase gene promoter, or the barley hordein gene promoter. Aroot-specific promoter can be the root specific subdomains of the CaMV35S promoter or the tobacco RD2 gene promoter. A non-seed fruit tissuepromoter can be a polygalacturonidase promoter, the banana TRX promoter,or the melon actin promoter. A promoter can be a constitutive promoter,such as 35S, p32449, or p13879. A promoter can be an inducible promoter.

Plant cells are also provided. In one embodiment, a plant cell caninclude an exogenous nucleic acid including a nucleic acid sequenceencoding a polypeptide having 80 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ IDNO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3,SEQ ID NO:8, and the Consensus sequence set forth in FIG. 6, whereexpression of the exogenous nucleic acid in a plant produced from theplant cell is effective to result in a different sugar level as comparedto a sugar level in a corresponding control plant that does not includethe exogenous nucleic acid. The sequence identity can be 85 percent orgreater, 90 percent or greater, or 95 percent or greater.

In another embodiment, a plant cell is provided. The plant cell caninclude an exogenous nucleic acid including a nucleic acid sequenceencoding a polypeptide having 80 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, (SEQ IDNO:11, SEQ ID NO:4, SEQ ID NO:9, and the Consensus sequence set forth inFIG. 6, where expression of the exogenous nucleic acid in a plantproduced from the plant cell is effective to result in a different sugarlevel as compared to a sugar level in a corresponding control plant thatdoes not include the exogenous nucleic acid. The sequence identity canbe 85 percent or greater, 90 percent or greater, or 95 percent orgreater.

In another aspect, a plant cell is provided. The plant cell can includean exogenous nucleic acid including a nucleic acid sequence encoding apolypeptide having 80 percent or greater sequence identity to an aminoacid sequence selected from the group consisting of SEQ ID NO:14, SEQ IDNO:2, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:4, and the Consensus sequenceset forth in FIG. 6, where expression of the exogenous nucleic acid in aplant produced from the plant cell is effective to result in a differentsugar level as compared to a sugar level in a corresponding controlplant that does not include the exogenous nucleic acid. The sequenceidentity can be 85 percent or greater, 90 percent or greater, or 95percent or greater.

In yet another embodiment, a plant cell is provided. The plant cell caninclude an exogenous nucleic acid including a nucleic acid sequenceencoding a polypeptide having 80 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:4, whereexpression of the exogenous nucleic acid in a plant produced from theplant cell is effective to result in a different sugar level as comparedto a sugar level in a corresponding control plant that does not includethe exogenous nucleic acid. The sequence identity can be 85 percent orgreater, 90 percent or greater, or 95 percent or greater.

In a further embodiment, a plant cell is provided. The plant cell caninclude an exogenous nucleic acid including a nucleic acid sequenceencoding a polypeptide including an amino acid sequence corresponding toSEQ ID NO:2, where expression of the exogenous nucleic acid in a plantproduced from the plant cell is effective to result in a different sugarlevel as compared to a sugar level in a corresponding control plant thatdoes not include the exogenous nucleic acid.

In another embodiment, a plant cell is provided. The plant cell caninclude an exogenous nucleic acid including a nucleic acid sequenceencoding a polypeptide including an amino acid sequence corresponding toSEQ ID NO:14, where expression of the exogenous nucleic acid in a plantproduced from the plant cell is effective to result in a different sugarlevel as compared to a sugar level in a corresponding control plant thatdoes not include the exogenous nucleic acid.

In another aspect, a plant cell is provided. The plant cell can includean exogenous nucleic acid including a nucleic acid sequence encoding apolypeptide including an amino acid sequence corresponding to theConsensus sequence set forth in FIG. 6, where expression of theexogenous nucleic acid in a plant produced from the plant cell iseffective to result in a different sugar level as compared to a sugarlevel in a corresponding control plant that does not include theexogenous nucleic acid.

In still another aspect, a plant cell is provided. The plant cell caninclude (a) a first exogenous nucleic acid including a nucleic acidsequence encoding a polypeptide having 80 percent or greater sequenceidentity to an amino acid sequence selected from the group consisting ofSEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ IDNO:8, and the Consensus sequence set forth in FIG. 6; and (b) a secondexogenous nucleic acid including a nucleic acid sequence encoding apolypeptide having 80 percent or greater sequence identity to an aminoacid sequence corresponding to SEQ ID NO:14; where expression of thefirst exogenous nucleic acid and the second exogenous nucleic acid in aplant produced from the plant cell is effective to result in a differentsugar level as compared to a sugar level in a corresponding controlplant that does not include the first exogenous nucleic acid or thesecond exogenous nucleic acid.

In another embodiment, a plant cell is provided. The plant cell caninclude (a) a first exogenous nucleic acid including a nucleic acidsequence encoding a polypeptide including an amino acid sequenceselected from the group consisting of SEQ ID NO:5, SEQ ID NO:10, SEQ IDNO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:2, SEQ ID NO:7,SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, and the Consensus sequence setforth in FIG. 6; and (b) a second exogenous nucleic acid including anucleic acid sequence encoding a polypeptide including an amino acidsequence corresponding to SEQ ID NO:14; where expression of the firstexogenous nucleic acid and the second exogenous nucleic acid in a plantproduced from the plant cell is effective to result in a different sugarlevel as compared to a sugar level in a corresponding control plant thatdoes not include the first exogenous nucleic acid or the secondexogenous nucleic acid.

In another embodiment, a plant cell is provided. The plant cell caninclude (a) a first exogenous nucleic acid including a nucleic acidsequence encoding a polypeptide having 80 percent or greater sequenceidentity to an amino acid sequence selected from the group consisting ofSEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, and SEQ IDNO:8; and (b) a second exogenous nucleic acid including a nucleic acidsequence encoding a polypeptide having 80 percent or greater sequenceidentity to an amino acid sequence corresponding to SEQ ID NO:14; whereexpression of the first exogenous nucleic acid and the second exogenousnucleic acid in a plant produced from the plant cell is effective toresult in a different sugar level as compared to a sugar level in acorresponding control plant that does not include the first exogenousnucleic acid or the second exogenous nucleic acid.

In yet another embodiment, a plant cell is provided. The plant cell caninclude (a) a first exogenous nucleic acid including a nucleic acidsequence encoding a polypeptide including an amino acid sequenceselected from the group consisting of SEQ ID NO:5, SEQ ID NO:10, SEQ IDNO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:2, SEQ ID NO:7,SEQ ID NO:12, SEQ ID NO:3, and SEQ ID NO:8; and (b) a second exogenousnucleic acid including a nucleic acid sequence encoding a polypeptideincluding an amino acid sequence corresponding to SEQ ID NO:14; whereexpression of the first exogenous nucleic acid and the second exogenousnucleic acid in a plant produced from the plant cell is effective toresult in a different sugar level as compared to a sugar level in acorresponding control plant that does not include the first exogenousnucleic acid or the second exogenous nucleic acid.

A different sugar level can be an increased level of one or more sugars,such as glucose, fructose, or sucrose. A different sugar level can be anincreased level of glucose and fructose, or an increased level ofglucose, fructose, and sucrose. An increased level of one or more sugarscan be in the non-seed tissue, seed, root, or fruit of a plant producedfrom a plant cell.

A plant or plant cell can be a member of one of the following genera:Abies, Agrostis, Allium, Alseodaphne, Anacardium, Ananus, Andropogon,Arabidopsis, Arachis, Apium, Aragrostis, Ascophyllum, Asparagus, Atropa,Avena, Beilschmiedia, Bixa, Brassica, Calendula, Capsicum, Carthamus,Chondrus, Chicorium, Cinnamomum, Citrus, Citrullus, Cocculus, Cocos,Coffea, Corylus, Cracilaria, Croton, Crypthecodinium, Cucumis,Cucurbita, Cunninghamia, Cuphea, Cynodon, Daucus, Dianthus, Duguetia,Elaeis, Enteromorpha, Euphoria, Festuca, Festulolium, Ficus, Fragaria,Fucus, Glaucium, Glycine, Glycyrrhiza, Gossypium, Haematococcus,Helianthus, Heterocallis, Hevea, Himanthalia, Hordeum, Hyoscyamus,Lactuca, Landolphia, Lemna, Linum, Litsea, Lolium, Lycopersicon,Lupinus, Majorana, Malus, Manihot, Medicago, Mentha, Musa, Nicotiana,Odontella, Olea, Oryza, Palmaria, Panicum, Pannesetum, Parthenium,Persea, Petunia, Phaseolus, Phleum, Phoenix, Picea, Pinus, Pistacia,Pisum, Poa, Populus sect., Porphyra, Prunus, Pyrus, Raphanus, Ricinus,Rosa, Rosmarinus, Rubus, Saccharum, Salix, Schizochytrium, Secale,Senecio, Sinapis, Solanum, Sorghum, Spinacia, Spirulina, Stephania,Triticum, Tagetes, Theobroma, Trifolium, Trigonella, Ulva, Undaria,Vaccinium, Vficia, Vigna, Vitis, Zea.

A plant or plant cell can be a member of one of the following species:Ananus comosus, Arabidopsis thaliana, Brassica rapa, Brassica napus,Brassica oleracea, Bixa orellana, Calendula officinalis, Cinnamomumcamphora, Coffea arabica, Glycine max, Glycyrrhiza glabra, Gossypiumhirsutum, Gossypium herbaceum, Lactuca sativa, Lycopersicon esculentum,Mentha piperita, Mentha spicata, Musa paradisiaca, Oryza sativa,Parthenium argentatum, Rosmarinus officinalis, Solanum tuberosum,Theobroma cacao, Triticum aestivum, Vitis vinifera, and Zea mays.

A plant or plant cell can be one of the following: alfalfa, amaranth,apple, beans (including kidney beans, lima beans, dry beans, greenbeans), broccoli, cabbage, carrot, castor bean, chick peas, cherry,chicory, chocolate, clover, coffee, cotton, cottonseed, crambe,eucalyptus, flax, grape, grapefruit, lemon, lentils, lettuce, linseed,mango, melon (e.g., watermelon, cantaloupe), mustard, orange, peanut,peach, pear, peas, pepper, plum, poplar, potato, rapeseed (high erucicacid and canola), safflower, sesame, soybean, spinach, strawberry,sugarbeet, sunflower, tea, tomato, banana, barley, date palm, fieldcorn, garlic, millet, oat, oil palm, onion, pineapple, popcorn, rice,rye, sorghum, sudangrass, sugarcane, sweet corn, switchgrass, turfgrasses, wheat, fir, pine, spruce, brown seaweeds, green seaweeds, redseaweeds, and microalgae.

Transgenic plants having modulated levels of one or more sugars ascompared to corresponding control plants are also provided. In oneembodiment, a transgenic plant can include an exogenous nucleic acidincluding a nucleic acid sequence encoding a polypeptide having 80percent or greater sequence identity to an amino acid sequence selectedfrom the group consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ IDNO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, and the Consensus sequenceset forth in FIG. 6. The sequence identity can be 85 percent or greater,90 percent or greater, or 95 percent or greater.

In another embodiment, a transgenic plant having a modulated level ofone or more sugars as compared to a corresponding control plant isprovided. The transgenic plant can include an exogenous nucleic acidincluding a nucleic acid sequence encoding a polypeptide having 80percent or greater sequence identity to an amino acid sequence selectedfrom the group consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, and theConsensus sequence set forth in FIG. 6. The sequence identity can be 85percent or greater, 90 percent or greater, or 95 percent or greater.

In another aspect, a transgenic plant having a modulated level of one ormore sugars as compared to a corresponding control plant is provided.The transgenic plant can include an exogenous nucleic acid including anucleic acid sequence encoding a polypeptide having 80 percent orgreater sequence identity to an amino acid sequence selected from thegroup consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6,SEQ ID NO:4, and the Consensus sequence set forth in FIG. 6. Thesequence identity can be 85 percent or greater, 90 percent or greater,or 95 percent or greater.

In yet another aspect, a transgenic plant having a modulated level ofone or more sugars as compared to a corresponding control plant isprovided. The transgenic plant can include an exogenous nucleic acidincluding a nucleic acid sequence encoding a polypeptide having 80percent or greater sequence identity to an amino acid sequence selectedfrom the group consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQID NO:6, and SEQ ID NO:4. The sequence identity can be 85 percent orgreater, 90 percent or greater, or 95 percent or greater.

In another embodiment, a transgenic plant having a modulated level ofone or more sugars as compared to a corresponding control plant isprovided. The transgenic plant can include an exogenous nucleic acidincluding a nucleic acid sequence encoding a polypeptide including anamino acid sequence corresponding to SEQ ID NO:2.

In a further embodiment, a transgenic plant having a modulated level ofone or more sugars as compared to a corresponding control plant isprovided. The transgenic plant can include an exogenous nucleic acidincluding a nucleic acid sequence encoding a polypeptide including anamino acid sequence corresponding to SEQ ID NO:14.

In another embodiment, a transgenic plant having a modulated level ofone or more sugars as compared to a corresponding control plant isprovided. The transgenic plant can include an exogenous nucleic acidincluding a nucleic acid sequence encoding a polypeptide including anamino acid sequence corresponding to the Consensus sequence set forth inFIG. 6.

In yet another embodiment, a transgenic plant having a modulated levelof one or more sugars as compared to a corresponding control plant isprovided. The transgenic plant can include (a) a first exogenous nucleicacid including a nucleic acid sequence encoding a polypeptide having 80percent or greater sequence identity to an amino acid sequence selectedfrom the group consisting of SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ IDNO:12, SEQ ID NO:3, SEQ ID NO:8, and the Consensus sequence set forth inFIG. 6; and (b) a second exogenous nucleic acid including a nucleic acidsequence encoding a polypeptide having 80 percent or greater sequenceidentity to an amino acid sequence corresponding to SEQ ID NO:14.

In another aspect, a transgenic plant having a modulated level of one ormore sugars as compared to a corresponding control plant is provided.The transgenic plant can include (a) a first exogenous nucleic acidincluding a nucleic acid sequence encoding a polypeptide including anamino acid sequence selected from the group consisting of SEQ ID NO:5,SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, and theConsensus sequence set forth in FIG. 6; and (b) a second exogenousnucleic acid including a nucleic acid sequence encoding a polypeptideincluding an amino acid sequence corresponding to SEQ ID NO:14.

In still a further aspect, a transgenic plant having a modulated levelof one or more sugars as compared to a corresponding control plant isprovided. The transgenic plant can include (a) a first exogenous nucleicacid including a nucleic acid sequence encoding a polypeptide having 80percent or greater sequence identity to an amino acid sequence selectedfrom the group consisting of SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ IDNO:12, SEQ ID NO:3, and SEQ ID NO:8; and (b) a second exogenous nucleicacid including a nucleic acid sequence encoding a polypeptide having 80percent or greater sequence identity to an amino acid sequencecorresponding to SEQ ID NO:14.

In yet another aspect, a transgenic plant having a modulated level ofone or more sugars as compared to a corresponding control plant isprovided. The transgenic plant can include (a) a first exogenous nucleicacid including a nucleic acid sequence encoding a polypeptide includingan amino acid sequence selected from the group consisting of SEQ IDNO:5, SEQ ID NO:I0, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9,SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, and SEQ ID NO:8;and (b) a second exogenous nucleic acid including a nucleic acidsequence encoding a polypeptide including an amino acid sequencecorresponding to SEQ ID NO:14.

Also provided are transgenic plant products, methods of producingproducts from transgenic plants, and articles of manufacture producedfrom transgenic plants. In one embodiment, tissues from a transgenicplant are provided, such as non-seed tissue, stalk, seed, or fruit. Inanother embodiment, a food product including non-seed tissue from atransgenic plant is provided. In a further embodiment, a food productincluding seed from a transgenic plant is provided. In anotherembodiment, animal feed including non-seed tissue or seeds is provided.In yet another embodiment, animal feed derived from a stalk is provided.

In one embodiment, a method of producing a sugar is provided. The methodincludes extracting a sugar from a transgenic plant provided herein,such as sugarcane or sugarbeet. The extract can be a liquid or a solid.The sugar can be one or more of sucrose, glucose, and/or fructose.

In another embodiment, a method of producing ethanol is provided. Themethod includes fermenting plant material from a transgenic plantprovided herein, such as corn.

In another aspect, articles of manufacture based on transgenic plantsare provided, including sugar, molasses, a bag of seeds, a bag of sugar,a bottle of sugar syrup, a liquid extract, or a solid extract.

An isolated nucleic acid is also provided. The isolated nucleic acid caninclude a nucleic acid sequence encoding a polypeptide having 80 percentor greater sequence identity to an amino acid sequence selected from thegroup consisting SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:10,SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:7, SEQ IDNO:12, SEQ ID NO:3, SEQ ID NO:8, and the Consensus sequence set forth inFIG. 6. A recombinant vector including the nucleic acid is alsoprovided.

Sugar-modulating polypeptides are provided herein. A sugar-modulatingpolypeptide can include the amino acid sequence corresponding to SEQ IDNO:5. A sugar-modulating polypeptide can include the amino acid sequencecorresponding to SEQ ID NO:10. A sugar-modulating polypeptide caninclude the amino acid sequence corresponding to SEQ ID NO:6. Asugar-modulating polypeptide can include the amino acid sequencecorresponding to SEQ ID NO:11. A sugar-modulating polypeptide caninclude the amino acid sequence corresponding to SEQ ID NO:4. Asugar-modulating polypeptide can include the amino acid sequencecorresponding to SEQ ID NO:9. A sugar-modulating polypeptide can includethe amino acid sequence corresponding to SEQ ID NO:7. A sugar-modulatingpolypeptide can include the amino acid sequence corresponding to SEQ IDNO:12. A sugar-modulating polypeptide can include the amino acidsequence corresponding to SEQ ID NO:3. A sugar-modulating polypeptidecan include the amino acid sequence corresponding to SEQ ID NO:8. Asugar-modulating polypeptide can include the amino acid sequencecorresponding to the Consensus sequence set forth in FIG. 6.

A sugar-modulating polypeptide can include a polypeptide having at least80 percent sequence identity (e.g., 80 percent, 85 percent, 90 percent,93 percent, 95 percent, 97 percent, 98 percent, or 99 percent sequenceidentity) to an amino acid sequence corresponding to SEQ ID NO:5. Asugar-modulating polypeptide can include a polypeptide having at least80 percent sequence identity (e.g., 80 percent, 85 percent, 90 percent,93 percent, 95 percent, 97 percent, 98 percent, or 99 percent sequenceidentity) to an amino acid sequence corresponding to SEQ ID NO:10. Asugar-modulating polypeptide can include a polypeptide having at least80 percent sequence identity (e.g., 80 percent, 85 percent, 90 percent,93 percent, 95 percent, 97 percent, 98 percent, or 99 percent sequenceidentity) to an amino acid sequence corresponding to SEQ ID NO:6. Asugar-modulating polypeptide can include a polypeptide having at least80 percent sequence identity (e.g., 80 percent, 85 percent, 90 percent,93 percent, 95 percent, 97 percent, 98 percent, or 99 percent sequenceidentity) to an amino acid sequence corresponding to SEQ ID NO:11. Insome cases, a sugar-modulating polypeptide can include a polypeptidehaving at least 80 percent sequence identity (e.g., 80 percent, 85percent, 90 percent, 93 percent, 95 percent, 97 percent, 98 percent, or99 percent sequence identity) to an amino acid sequence corresponding toSEQ ID NO:4. A sugar-modulating polypeptide can include a polypeptidehaving at least 80 percent sequence identity (e.g., 80 percent, 85percent, 90 percent, 93 percent, 95 percent, 97 percent, 98 percent, or99 percent sequence identity) to an amino acid sequence corresponding toSEQ ID NO:9. In some cases, a sugar-modulating polypeptide can include apolypeptide having at least 80 percent sequence identity (e.g., 80percent, 85 percent, 90 percent, 93 percent, 95 percent, 97 percent, 98percent, or 99 percent sequence identity) to an amino acid sequencecorresponding to SEQ ID NO:7. A sugar-modulating polypeptide can includea polypeptide having at least 80 percent sequence identity (e.g., 80percent, 85 percent, 90 percent, 93 percent, 95 percent, 97 percent, 98percent, or 99 percent sequence identity) to an amino acid sequencecorresponding to SEQ ID NO:12. A sugar-modulating polypeptide caninclude a polypeptide having at least 80 percent sequence identity(e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95 percent, 97percent, 98 percent, or 99 percent sequence identity) to an amino acidsequence corresponding to SEQ ID NO:3. A sugar-modulating polypeptidecan include a polypeptide having at least 80 percent sequence identity(e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95 percent, 97percent, 98 percent, or 99 percent sequence identity) to an amino acidsequence corresponding to SEQ ID NO:8. A sugar-modulating polypeptidecan include a polypeptide having at least 80 percent sequence identity(e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95 percent, 97percent, 98 percent, or 99 percent sequence identity) to an amino acidsequence corresponding to the Consensus sequence set forth in FIG. 6.

Nucleic acids encoding sugar-modulating polypeptides are providedherein. Such nucleic acids can be used to transform plant cells. Anucleic acid encoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:5 can be used to transform a plant cell. Anucleic acid encoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:10 can be used to transform a plant cell. Anucleic acid encoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:6 can be used to transform a plant cell. Anucleic acid encoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:11 can be used to transform a plant cell. Anucleic acid encoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:4 can be used to transform a plant cell. Anucleic acid encoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:9 can be used to transform a plant cell. Anucleic acid encoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:7 can be used to transform a plant cell. Anucleic acid encoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:12 can be used to transform a plant cell. Anucleic acid encoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:3 can be used to transform a plant cell. Anucleic acid encoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:8 can be used to transform a plant cell. Anucleic acid encoding a polypeptide that includes an amino acid sequencecorresponding to the Consensus sequence set forth in FIG. 6 can be usedto transform a plant cell.

A nucleic acid encoding a polypeptide having at least 80 percentsequence identity (e.g., 80 percent, 85 percent, 90 percent, 93 percent,95 percent, 97 percent, 98 percent, or 99 percent sequence identity) toan amino acid sequence corresponding to SEQ ID NO:5 can be used totransform a plant cell. In some cases, a nucleic acid encoding apolypeptide having at least 80 percent sequence identity (e.g., 80percent, 85 percent, 90 percent, 93 percent, 95 percent, 97 percent, 98percent, or 99 percent sequence identity) to an amino acid sequencecorresponding to SEQ ID NO:10 can be used to transform a plant cell. Anucleic acid encoding a polypeptide having at least 80 percent sequenceidentity (e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95percent, 97 percent, 98 percent, or 99 percent sequence identity) to anamino acid sequence corresponding to SEQ ID NO:6 can be used totransform a plant cell. A nucleic acid encoding a polypeptide having atleast 80 percent sequence identity (e.g., 80 percent, 85 percent, 90percent, 93 percent, 95 percent, 97 percent, 98 percent, or 99 percentsequence identity) to an amino acid sequence corresponding to SEQ IDNO:11 can be used to transform a plant cell. A nucleic acid encoding apolypeptide having at least 80 percent sequence identity (e.g., 80percent, 85 percent, 90 percent, 93 percent, 95 percent, 97 percent, 98percent, or 99 percent sequence identity) to an amino acid sequencecorresponding to SEQ ID NO:4 can be used to transform a plant cell. Insome cases, a nucleic acid encoding a polypeptide having at least 80percent sequence identity (e.g., 80 percent, 85 percent, 90 percent, 93percent, 95 percent, 97 percent, 98 percent, or 99 percent sequenceidentity) to an amino acid sequence corresponding to SEQ ID NO:9 can beused to transform a plant cell. A nucleic acid encoding a polypeptidehaving at least 80 percent sequence identity (e.g., 80 percent, 85percent, 90 percent, 93 percent, 95 percent, 97 percent, 98 percent, or99 percent sequence identity) to an amino acid sequence corresponding toSEQ ID NO:7 can be used to transform a plant cell. A nucleic acidencoding a polypeptide having at least 80 percent sequence identity(e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95 percent, 97percent, 98 percent, or 99 percent sequence identity) to an amino acidsequence corresponding to SEQ ID NO:12 can be used to transform a plantcell. A nucleic acid encoding a polypeptide having at least 80 percentsequence identity (e.g., 80 percent, 85 percent, 90 percent, 93 percent,95 percent, 97 percent, 98 percent, or 99 percent sequence identity) toan amino acid sequence corresponding to SEQ ID NO:3 can be used totransform a plant cell. A nucleic acid encoding a polypeptide having atleast 80 percent sequence identity (e.g., 80 percent, 85 percent, 90percent, 93 percent, 95 percent, 97 percent, 98 percent, or 99 percentsequence identity) to an amino acid sequence corresponding to SEQ IDNO:8 can be used to transform a plant cell. In some cases, a nucleicacid encoding a polypeptide having at least 80 percent sequence identity(e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95 percent, 97percent, 98 percent, or 99 percent sequence identity) to an amino acidsequence corresponding to the Consensus sequence set forth in FIG. 6 canbe used to transform a plant cell.

A first nucleic acid encoding a polypeptide that includes an amino acidsequence corresponding to SEQ ID NO:5, and a second nucleic acidencoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:14 can be used to transform a plant cell. Afirst nucleic acid encoding a polypeptide that includes an amino acidsequence corresponding to SEQ ID NO:10, and a second nucleic acidencoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:14 can be used to transform a plant cell. Insome cases, a first nucleic acid encoding a polypeptide that includes anamino acid sequence corresponding to SEQ ID NO:6, and a second nucleicacid encoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:14 can be used to transform a plant cell. Afirst nucleic acid encoding a polypeptide that includes an amino acidsequence corresponding to SEQ ID NO:11, and a second nucleic acidencoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:14 can be used to transform a plant cell. Afirst nucleic acid encoding a polypeptide that includes an amino acidsequence corresponding to SEQ ID NO:4, and a second nucleic acidencoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:14 can be used to transform a plant cell. Afirst nucleic acid encoding a polypeptide that includes an amino acidsequence corresponding to SEQ ID NO:9, and a second nucleic acidencoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:14 can be used to transform a plant cell. Insome cases, a first nucleic acid encoding a polypeptide that includes anamino acid sequence corresponding to SEQ ID NO:7, and a second nucleicacid encoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:14 can be used to transform a plant cell. Afirst nucleic acid encoding a polypeptide that includes an amino acidsequence corresponding to SEQ ID NO:12, and a second nucleic acidencoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:14 can be used to transform a plant cell. Afirst nucleic acid encoding a polypeptide that includes an amino acidsequence corresponding to SEQ ID NO:3, and a second nucleic acidencoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:14 can be used to transform a plant cell. Insome cases, a first nucleic acid encoding a polypeptide that includes anamino acid sequence corresponding to SEQ ID NO:8, and a second nucleicacid encoding a polypeptide that includes an amino acid sequencecorresponding to SEQ ID NO:14 can be used to transform a plant cell. Afirst nucleic acid encoding a polypeptide that includes an amino acidsequence corresponding to the Consensus sequence set forth in FIG. 6,and a second nucleic acid encoding a polypeptide that includes an aminoacid sequence corresponding to SEQ ID NO:14 can be used to transform aplant cell.

A first nucleic acid encoding a polypeptide having at least 80 percentsequence identity (e.g., 80 percent, 85 percent, 90 percent, 93 percent,95 percent, 97 percent, 98 percent, or 99 percent sequence identity) toan amino acid sequence corresponding to SEQ ID NO:5, and a secondnucleic acid encoding a polypeptide having at least 80 percent sequenceidentity (e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95percent, 97 percent, 98 percent, or 99 percent sequence identity) to anamino acid sequence corresponding to SEQ ID NO:14 can be used totransform a plant cell. A first nucleic acid encoding a polypeptidehaving at least 80 percent sequence identity (e.g., 80 percent, 85percent, 90 percent, 93 percent, 95 percent, 97 percent, 98 percent, or99 percent sequence identity) to an amino acid sequence corresponding toSEQ ID NO:10, and a second nucleic acid encoding a polypeptide having atleast 80 percent sequence identity (e.g., 80 percent, 85 percent, 90percent, 93 percent, 95 percent, 97 percent, 98 percent, or 99 percentsequence identity) to an amino acid sequence corresponding to SEQ IDNO:14 can be used to transform a plant cell. A first nucleic acidencoding a polypeptide having at least 80 percent sequence identity(e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95 percent, 97percent, 98 percent, or 99 percent sequence identity) to an amino acidsequence corresponding to SEQ ID NO:6, and a second nucleic acidencoding a polypeptide having at least 80 percent sequence identity(e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95 percent, 97percent, 98 percent, or 99 percent sequence identity) to an amino acidsequence corresponding to SEQ ID NO:14 can be used to transform a plantcell. A first nucleic acid encoding a polypeptide having at least 80percent sequence identity (e.g., 80 percent, 85 percent, 90 percent, 93percent, 95 percent, 97 percent, 98 percent, or 99 percent sequenceidentity) to an amino acid sequence corresponding to SEQ ID NO:11, and asecond nucleic acid encoding a polypeptide having at least 80 percentsequence identity (e.g., 80 percent, 85 percent, 90 percent, 93 percent,95 percent, 97 percent, 98 percent, or 99 percent sequence identity) toan amino acid sequence corresponding to SEQ ID NO:14 can be used totransform a plant cell. A first nucleic acid encoding a polypeptidehaving at least 80 percent sequence identity (e.g., 80 percent, 85percent, 90 percent, 93 percent, 95 percent, 97 percent, 98 percent, or99 percent sequence identity) to an amino acid sequence corresponding toSEQ ID NO:4, and a second nucleic acid encoding a polypeptide having atleast 80 percent sequence identity (e.g., 80 percent, 85 percent, 90percent, 93 percent, 95 percent, 97 percent, 98 percent, or 99 percentsequence identity) to an amino acid sequence corresponding to SEQ IDNO:14 can be used to transform a plant cell. A first nucleic acidencoding a polypeptide having at least 80 percent sequence identity(e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95 percent, 97percent, 98 percent, or 99 percent sequence identity) to an amino acidsequence corresponding to SEQ ID NO:9, and a second nucleic acidencoding a polypeptide having at least 80 percent sequence identity(e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95 percent, 97percent, 98 percent, or 99 percent sequence identity) to an amino acidsequence corresponding to SEQ ID NO:14 can be used to transform a plantcell. A first nucleic acid encoding a polypeptide having at least 80percent sequence identity (e.g., 80 percent, 85 percent, 90 percent, 93percent, 95 percent, 97 percent, 98 percent, or 99 percent sequenceidentity) to an amino acid sequence corresponding to SEQ ID NO:7, and asecond nucleic acid encoding a polypeptide having at least 80 percentsequence identity (e.g., 80 percent, 85 percent, 90 percent, 93 percent,95 percent, 97 percent, 98 percent, or 99 percent sequence identity) toan amino acid sequence corresponding to SEQ ID NO:14 can be used totransform a plant cell. A first nucleic acid encoding a polypeptidehaving at least 80 percent sequence identity (e.g., 80 percent, 85percent, 90 percent, 93 percent, 95 percent, 97 percent, 98 percent, or99 percent sequence identity) to an amino acid sequence corresponding toSEQ ID NO:12, and a second nucleic acid encoding a polypeptide having atleast 80 percent sequence identity (e.g., 80 percent, 85 percent, 90percent, 93 percent, 95 percent, 97 percent, 98 percent, or 99 percentsequence identity) to an amino acid sequence corresponding to SEQ IDNO:14 can be used to transform a plant cell. A first nucleic acidencoding a polypeptide having at least 80 percent sequence identity(e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95 percent, 97percent, 98 percent, or 99 percent sequence identity) to an amino acidsequence corresponding to SEQ ID NO:3, and a second nucleic acidencoding a polypeptide having at least 80 percent sequence identity(e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95 percent, 97percent, 98 percent, or 99 percent sequence identity) to an amino acidsequence corresponding to SEQ ID NO:14 can be used to transform a plantcell. A first nucleic acid encoding a polypeptide having at least 80percent sequence identity (e.g., 80 percent, 85 percent, 90 percent, 93percent, 95 percent, 97 percent, 98 percent, or 99 percent sequenceidentity) to an amino acid sequence corresponding to SEQ ID NO:8, and asecond nucleic acid encoding a polypeptide having at least 80 percentsequence identity (e.g., 80 percent, 85 percent, 90 percent, 93 percent,95 percent, 97 percent, 98 percent, or 99 percent sequence identity) toan amino acid sequence corresponding to SEQ ID NO:14 can be used totransform a plant cell. A first nucleic acid encoding a polypeptidehaving at least 80 percent sequence identity (e.g., 80 percent, 85percent, 90 percent, 93 percent, 95 percent, 97 percent, 98 percent, or99 percent sequence identity) to an amino acid sequence corresponding tothe Consensus sequence set forth in FIG. 6, and a second nucleic acidencoding a polypeptide having at least 80 percent sequence identity(e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95 percent, 97percent, 98 percent, or 99 percent sequence identity) to an amino acidsequence corresponding to SEQ ID NO:14 can be used to transform a plantcell.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention pertains. Although methods and materialssimilar or equivalent to those described herein can be used to practicethe invention, suitable methods and materials are described below. Allpublications, patent applications, patents, and other referencesmentioned herein are incorporated by reference in their entirety. Incase of conflict, the present specification, including definitions, willcontrol. In addition, the materials, methods, and examples areillustrative only and not intended to be limiting.

The details of one or more embodiments of the invention are set forth inthe accompanying drawings and the description below. Other features,objects, and advantages of the invention will be apparent from thedescription and drawings, and from the claims.

DESCRIPTION OF THE DRAWINGS

FIG. 1 is the nucleotide sequence of Ceres clone 625627 (SEQ ID NO:13).

FIG. 2 is the amino acid sequence encoded by Ceres clone 625627 (SEQ IDNO:14).

FIG. 3 is the nucleotide sequence of Ceres clone 32380 (SEQ ID NO:1).

FIG. 4 is the amino acid sequence encoded by Ceres clone 32380 (SEQ IDNO:2).

FIG. 5 is a graph plotting levels of glucose and fructose in T₂ ME02225plants relative to control plants.

FIG. 6 is an alignment of the amino acid sequence of SEQ ID NO:2 withfunctionally homologous and orthologous amino acid sequences SEQ IDNO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:7, SEQ IDNO:12, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:3, and SEQ ID NO:8. Aconsensus sequence determined by the alignment is set forth.

DETAILED DESCRIPTION

The materials and methods provided herein can be used to make plants,plant tissues, and plant products having modulated levels of sugars(e.g., glucose, fructose, and sucrose). For example, plants havingincreased levels of sugars in seeds and/or non-seed tissues are providedherein. The methods can include transforming a plant cell with one ormore nucleic acids that encode sugar-modulating polypeptides, whereinexpression of the one or more polypeptides results in modulated levels(e.g., increased or decreased levels) of one or more sugars. Plants andplant materials (e.g., seeds, non-seed tissues) produced using suchmethods can be used as food sources of sugars, or as sources of sugarsfor inclusion in nutritional supplements, for example.

Polypeptides

Isolated polypeptides, including sugar-modulating polypeptides, areprovided herein. The term “polypeptide” as used herein refers to acompound of two or more subunit amino acids, amino acid analogs, orother peptidomimetics, regardless of post-translational modification(e.g., phosphorylation or glycosylation). The subunits may be linked bypeptide bonds or other bonds such as, for example, ester or ether bonds.The term “amino acid” refers to natural and/or unnatural or syntheticamino acids, including D/L optical isomers. Full-length proteins,analogs, mutants, and fragments thereof are encompassed by thisdefinition.

By “isolated” or “purified” with respect to a polypeptide it is meantthat the polypeptide is separated to some extent from the cellularcomponents with which it is normally found in nature (e.g, otherpolypeptides, lipids, carbohydrates, and nucleic acids). A purifiedpolypeptide can yield a single major band on a non-reducingpolyacrylamide gel. A purified polypeptide can be at least about 75percent pure (e.g., at least 80 percent, 85 percent, 90 percent, 95percent, 97 percent, 98 percent, 99 percent, or 100 percent pure).Purified polypeptides can be obtained by, for example, extraction from anatural source, by chemical synthesis, or by recombinant production in ahost cell or transgenic plant, and can be purified using, for example,affinity chromatography, immunoprecipitation, size exclusionchromatography, and ion exchange chromatography. The extent ofpurification can be measured using any appropriate method, including,without limitation, column chromatography, polyacrylamide gelelectrophoresis, or high-performance liquid chromatography.

Described herein are sugar-modulating polypeptides. A sugar-modulatingpolypeptide can be effective to modulate a level of one or more sugarsby any mechanism. For example, a sugar-modulating polypeptide canmodulate sugar biosynthesis, stability, and/or degradation. In somecases, such a polypeptide is a transcription factor containing an AP2DNA-binding domain. An AP2 DNA-binding domain is a distinguishingcharacteristic of a family of transcription factors unique to plants.The prototypic members of the family are AP2 (APETALA2) and EREBPs(ethylene-responsive element binding proteins). AP2/REBP genes form alarge multigene family, and they play a variety of roles throughout theplant life cycle: from being key regulators of several developmentalprocesses, like floral organ identity determination or control of leafepidermal cell identity, to forming part of the mechanisms used byplants to respond to various types of biotic and environmental stress.

SEQ ID NO:14 shown in FIG. 2 sets forth the amino acid sequence of aclone identified herein as Ceres clone 625627, which is predicted toinclude an AP2 DNA-binding domain. A sugar-modulating polypeptide can bea polypeptide including the amino acid sequence set forth in SEQ IDNO:14. Alternatively, a sugar-modulating polypeptide can be an ortholog,homolog, or variant of the polypeptide having the sequence set forth inSEQ ID NO:14. For example, a sugar-modulating polypeptide can have anamino acid sequence with at least 60 percent sequence identity (e.g., 61percent, 66 percent, 68 percent, 70 percent, 72 percent, 74 percent, 76percent, 78 percent, 80 percent, 81 percent, 82 percent, 83 percent, 84percent, 85 percent, 86 percent, 87 percent, 88 percent, 89 percent, 90percent, 91 percent, 92 percent, 93 percent, 94 percent, 95 percent, 96percent, 97 percent, 98 percent, or 99 percent sequence identity) to theamino acid sequence set forth in SEQ ID NO:14.

In other cases, a sugar-modulating polypeptide is a DNA-directed RNApolymerase, such as DNA-directed RNA polymerase II. A DNA-directed RNApolymerase catalyzes the transcription of DNA into RNA.

SEQ ID NO:2 shown in FIG. 4 sets forth the amino acid sequence of anArabidopsis clone identified herein as Ceres clone 32380, which ispredicted to include a DNA-directed RNA polymerase II third largestsubunit. Homologs and orthologs of the polypeptide having the amino acidsequence set forth in SEQ ID NO:2 are provided in FIG. 6.

A sugar-modulating polypeptide can be a polypeptide including the aminoacid sequence set forth in SEQ ID NO:2. Alternatively, asugar-modulating polypeptide can be an ortholog, homolog, or variant ofthe polypeptide having the sequence set forth in SEQ ID NO:2. Forexample, a sugar-modulating polypeptide can have an amino acid sequencewith at least 60 percent sequence identity (e.g., 61 percent, 66percent, 68 percent, 70 percent, 72 percent, 74 percent, 76 percent, 78percent, 80 percent, 81 percent, 82 percent, 83 percent, 84 percent, 85percent, 86 percent, 87 percent, 88 percent, 89 percent, 90 percent, 91percent, 92 percent, 93 percent, 94 percent, 95 percent, 96 percent, 97percent, 98 percent, or 99 percent sequence identity) to the amino acidsequence set forth in SEQ ID NO:2. For example, a sugar-modulatingpolypeptide can include the amino acid sequence corresponding to Ceresclone 698259 (SEQ ID NO:5), Ceres clone 698259T (SEQ ID NO:10), Ceresclone 244359 (SEQ ID NO:6), Ceres clone 244359T (SEQ ID NO:11), Ceresclone 692249 (SEQ ID NO:4), Ceres clone 692249T (SEQ ID NO:9),gi|50898416 (SEQ ID NO:7), gi|50898416T (SEQ ID NO:12), gi|21593370 (SEQID NO:3), gi|21593370T (SEQ ID NO:8), or the Consensus sequence setforth in FIG. 6.

In some cases, a sugar-modulating polypeptide can include a polypeptidehaving at least 80 percent sequence identity (e.g., 80 percent, 85percent, 90 percent, 93 percent, 95 percent, 97 percent, 98 percent, or99 percent sequence identity) to an amino acid sequence corresponding toSEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQID NO:9, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, or theConsensus sequence set forth in FIG. 6.

A consensus amino acid sequence for a sugar-modulating polypeptide canbe determined by aligning homologous and/or orthologous amino acidsequences (e.g., amino acid sequences set forth in FIG. 6) anddetermining the most common amino acid or type of amino acid at eachposition. For example, a consensus sequence can be determined byaligning amino acid sequences corresponding to SEQ ID NO:5, SEQ IDNO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:7, SEQ ID NO:12, SEQ IDNO:4, SEQ ID NO:9, SEQ ID NO:2, SEQ ID NO:3, and SEQ ID NO:8 as shown inFIG. 6.

Other means by which sugar-modulating polypeptides can be identifiedinclude functional complementation of sugar-modulating polypeptidemutants. Suitable sugar-modulating polypeptides also can be identifiedby analysis of nucleotide and polypeptide sequence alignments. Forexample, performing a query on a database of nucleotide or polypeptidesequences can identify homologs and/or orthologs of the polypeptidehaving the amino acid sequence set forth in SEQ ID NO:2. Sequenceanalysis can involve BLAST, Reciprocal BLAST, or PSI-BLAST analysis ofnonredundant databases. Those proteins in the database that have greaterthan 35% sequence identity to the specific query polypeptide can becandidates for further evaluation for suitability as sugar-modulatingpolypeptides. If desired, manual inspection of such candidates can becarried out in order to reduce the number of candidates to be furtherevaluated. Manual inspection can be performed by selecting thosecandidates that appear to have domains suspected of being present insugar-modulating polypeptides.

Typically, conserved regions of sugar-modulating polypeptides exhibit atleast 40 percent amino acid sequence identity (e.g., at least 45percent, at least 50 percent, at least 60 percent, at least 70 percent,at least 80 percent, or at least 90 percent amino acid sequenceidentity). Conserved regions of target and template polypeptides canexhibit at least 92 percent, 94 percent, 96 percent, 98 percent, or 99percent amino acid sequence identity. Amino acid sequence identity canbe deduced from amino acid or nucleotide sequences. In certain cases,highly conserved domains can be identified within sugar-modulatingpolypeptides. These conserved regions can be useful in identifyingfunctionally similar polypeptides.

Domains are groups of contiguous amino acids in a polypeptide that canbe used to characterize protein families and/or parts of proteins. Suchdomains have a “fingerprint” or “signature” that can comprise conserved(1) primary sequence, (2) secondary structure, and/or (3)three-dimensional conformation. Generally, each domain has beenassociated with either a conserved primary sequence or a sequence motif.Generally these conserved primary sequence motifs have been correlatedwith specific in vitro and/or in vivo activities. A domain can be anylength, including the entirety of the polynucleotide to be transcribed.

The identification of conserved regions in a template, or subject,polypeptide can facilitate production of variants of wild typesugar-modulating polypeptides. Conserved regions can be identified bylocating a region within the primary amino acid sequence of a templatepolypeptide that is a repeated sequence, forms some secondary structure(e.g., helices and beta sheets), establishes positively or negativelycharged domains, or represents a protein motif or domain. See, e.g., thePfam web site describing consensus sequences for a variety of proteinmotifs and domains on the World Wide Web at sanger.ac.uk/Pfam/ andonline at genome.wustl.edu/Pfam/. Descriptions of the informationincluded at the Pfam database are included in Sonnhammer et al., 1998,Nucl. Acids Res. 26:320-322; Sonnhammer et al., 1997, Proteins28:405-420; and Bateman et al., 1999, Nucl. Acids Res. 27:260-262. Fromthe Pfam database, consensus sequences of protein motifs and domains canbe aligned with the template polypeptide sequence to determine conservedregion(s).

Conserved regions also can be determined by aligning sequences of thesame or related polypeptides from closely related species. Closelyrelated species preferably are from the same family. In someembodiments, alignment of sequences from two different species isadequate. For example, sequences from Arabidopsis and Zea mays can beused to identify one or more conserved regions.

If desired, the classification of a polypeptide as a sugar-modulatingpolypeptide can be determined using techniques known to those havingordinary skill in the art. These techniques can be divided into twogeneral categories: global sugar analysis, and type-specific sugaranalysis. Global sugar analysis techniques can include determining theoverall level of sugars within a cell, group of cells, or tissue (e.g.,non-seed tissue vs. seed tissue). Type-specific sugar analysistechniques can include measuring the level of a particular type of sugar(i.e., glucose, fructose, or sucrose).

Polynucleotides

Isolated nucleic acids are also provided herein, including isolatednucleic acids that encode any of the sugar-modulating polypeptidesdescribed herein. The terms “nucleic acid” and “polynucleotide” are usedinterchangeably herein, and refer to both RNA and DNA, including cDNA,genomic DNA, synthetic (e.g., chemically synthesized) DNA, and DNA (orRNA) containing nucleic acid analogs. Polynucleotides can have anythree-dimensional structure. A nucleic acid can be double-stranded orsingle-stranded (i.e., a sense strand or an antisense strand).Non-limiting examples of polynucleotides include genes, gene fragments,exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA,siRNA, micro-RNA, ribozymes, cDNA, recombinant polynucleotides, branchedpolynucleotides, plasmids, vectors, isolated DNA of any sequence,isolated RNA of any sequence, nucleic acid probes, and primers, as wellas nucleic acid analogs.

As used herein, “isolated,” when in reference to a nucleic acid, refersto a nucleic acid that is separated from other nucleic acids that arepresent in a genome, e.g., a plant genome, including nucleic acids thatnormally flank one or both sides of the nucleic acid in the genome. Theterm “isolated” as used herein with respect to nucleic acids alsoincludes any non-naturally-occurring sequence, since suchnon-naturally-occurring sequences are not found in nature and do nothave immediately contiguous sequences in a naturally-occurring genome.

An isolated nucleic acid can be, for example, a DNA molecule, providedone of the nucleic acid sequences normally found immediately flankingthat DNA molecule in a naturally-occurring genome is removed or absent.Thus, an isolated nucleic acid includes, without limitation, a DNAmolecule that exists as a separate molecule (e.g., a chemicallysynthesized nucleic acid, or a cDNA or genomic DNA fragment produced bythe polymerase chain reaction (PCR) or restriction endonucleasetreatment) independent of other sequences, as well as DNA that isincorporated into a vector, an autonomously replicating plasmid, a virus(e.g., pararetrovirus, retrovirus, lentivirus, adenovirus,adeno-associated virus, or herpesvirus), or the purified genomic DNA ofa prokaryote or eukaryote. In addition, an isolated nucleic acid caninclude an engineered nucleic acid such as a DNA molecule that is partof a hybrid or fusion nucleic acid. A nucleic acid existing amonghundreds to millions of other nucleic acids within, for example, cDNAlibraries or genomic libraries, or gel slices containing a genomic DNArestriction digest, is not to be considered an isolated nucleic acid.

A nucleic acid can be made, for example, by chemical synthesis or usingPCR. PCR refers to a procedure or technique in which target nucleicacids are amplified. PCR can be used to amplify specific sequences fromDNA as well as RNA, including sequences from total genomic DNA or totalcellular RNA. Various PCR methods are described, for example, in PCRPrimer: A Laboratory Manual, Dieffenbach and Dveksler, eds., Cold SpringHarbor Laboratory Press, 1995. Generally, sequence information from theends of the region of interest or beyond is employed to designoligonucleotide primers that are identical or similar in sequence toopposite strands of the template to be amplified. Various PCR strategiesalso are available by which site-specific nucleotide sequencemodifications can be introduced into a template nucleic acid.

The term “exogenous” with respect to a nucleic acid indicates that thenucleic acid is part of a recombinant nucleic acid construct, or is notin its natural environment. For example, an exogenous nucleic acid canbe a sequence from one species introduced into another species, i.e., aheterologous nucleic acid. Typically, such an exogenous nucleic acid isintroduced into the other species via a recombinant nucleic acidconstruct. An exogenous nucleic acid can also be a sequence that isnative to an organism and that has been reintroduced into cells of thatorganism. An exogenous nucleic acid that includes a native sequence canoften be distinguished from the naturally occurring sequence by thepresence of non-natural sequences linked to the exogenous nucleic acid,e.g., non-native regulatory regions flanking a native sequence in arecombinant nucleic acid construct. In addition, stably transformedexogenous nucleic acids typically are integrated at positions other thanthe position where the native sequence is found. It will be appreciatedthat an exogenous nucleic acid may have been introduced into aprogenitor and not into the cell under consideration. For example, atransgenic plant containing an exogenous nucleic acid can be the progenyof a cross between a stably transformed plant and a non-transgenicplant. Such progeny are considered to contain the exogenous nucleicacid.

Thus, provided herein are nucleic acids encoding any of thesugar-modulating polypeptides described previously One example of anisolated polynucleotide is SEQ ID NO:13 shown in FIG. 1, which setsforth the nucleotide sequence of a clone identified herein as Ceresclone 625627. Another example of an isolated polynucleotide is SEQ IDNO:1 shown in FIG. 3, which sets forth the nucleotide sequence of anArabidopsis clone identified herein as Ceres clone 32380. Fragments,fusions, complements, and reverse complements of the describedpolynucleotides (and encoded polypeptides) also are contemplated.

One or more nucleic acids that encode sugar-modulating polypeptides canbe used to transform a plant cell such that a plant produced from theplant cell has a modulated (e.g., increased) level of one or moresugars. For example, a nucleic acid encoding a polypeptide that includesan amino acid sequence corresponding to SEQ ID NO:14 can be used totransform a plant cell. A nucleic acid encoding a polypeptide having atleast 80 percent sequence identity (e.g., 80 percent, 85 percent, 90percent, 93 percent, 95 percent, 97 percent, 98 percent, or 99 percentsequence identity) to an amino acid sequence corresponding to SEQ IDNO:14 can also be used to transform a plant cell.

In certain cases, a nucleic acid encoding a polypeptide that includes anamino acid sequence corresponding to SEQ ID NO:2 can be used totransform a plant cell. In other cases, a nucleic acid encoding apolypeptide that includes an amino acid sequence corresponding to SEQ IDNO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9,SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, or the Consensussequence set forth in FIG. 6 can be used to transform a plant cell.

In some cases, a nucleic acid encoding a polypeptide having at least 80percent sequence identity (e.g., 80 percent, 85 percent, 90 percent, 93percent, 95 percent, 97 percent, 98 percent, or 99 percent sequenceidentity) to an amino acid sequence corresponding to SEQ ID NO:2 can beused to transform a plant cell. In yet other cases, a nucleic acidencoding a polypeptide having at least 80 percent sequence identity(e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95 percent, 97percent, 98 percent, or 99 percent sequence identity) to an amino acidsequence corresponding to SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ IDNO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3,SEQ ID NO:8, or the Consensus sequence set forth in FIG. 6 can be usedto transform a plant cell.

Two or more nucleic acids that encode sugar-modulating polypeptides canalso be used to transform a plant cell such that a plant produced fromthe plant cell has a modulated (e.g., increased) level of one or moresugars. For example, a first nucleic acid encoding a polypeptide thatincludes an amino acid sequence corresponding to SEQ ID NO:2, and asecond nucleic acid encoding a polypeptide that includes an amino acidsequence corresponding to SEQ ID NO:14 can be used to transform a plantcell. In certain embodiments, a first nucleic acid encoding apolypeptide that includes an amino acid sequence corresponding to SEQ IDNO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9,SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, or the Consensussequence set forth in FIG. 6, and a second nucleic acid encoding apolypeptide that includes an amino acid sequence corresponding to SEQ IDNO:14 can be used to transform a plant cell.

In yet other cases, a first nucleic acid encoding a polypeptide havingat least 80 percent sequence identity (e.g., 80 percent, 85 percent, 90percent, 93 percent, 95 percent, 97 percent, 98 percent, or 99 percentsequence identity) to an amino acid sequence corresponding to SEQ IDNO:2, and a second nucleic acid encoding a polypeptide having at least80 percent sequence identity (e.g., 80 percent, 85 percent, 90 percent,93 percent, 95. percent, 97 percent, 98 percent, or 99 percent sequenceidentity) to an amino acid sequence corresponding to SEQ ID NO:14 can beused to transform a plant cell. In addition, a first nucleic acidencoding a polypeptide having at least 80 percent sequence identity(e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95 percent, 97percent, 98 percent, or 99 percent sequence identity) to an amino acidsequence corresponding to SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ IDNO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3,SEQ ID NO:8, or the Consensus sequence set forth in FIG. 6, and a secondnucleic acid encoding a polypeptide having at least 80 percent sequenceidentity (e.g., 80 percent, 85 percent, 90 percent, 93 percent, 95percent, 97 percent, 98 percent, or 99 percent sequence identity) to anamino acid sequence corresponding to SEQ ID NO:14 can be used totransform a plant cell.

It will be appreciated that methods described herein can utilizenon-transgenic plant cells or plants that carry a mutation in asugar-modulating polypeptide. For example, a plant carrying a T-DNAinsertion, a deletion, a transversion mutation, or a transition mutationin the coding sequence for one of the aforementioned polypeptides canaffect sugar levels.

As used herein, the term “percent sequence identity” refers to thedegree of identity between any given query sequence and a subjectsequence. A percent identity for any query nucleic acid or amino acidsequence, e.g., a sugar-modulating polypeptide, relative to anothersubject nucleic acid or amino acid sequence can be determined asfollows. A query nucleic acid or amino acid sequence is aligned to oneor more subject nucleic acid or amino acid sequences using the computerprogram ClustalW (version 1.83, default parameters), which allowsalignments of nucleic acid or protein sequences to be carried out acrosstheir entire length (global alignment).

ClustalW calculates the best match between a query and one or moresubject sequences, and aligns them so that identities, similarities anddifferences can be determined. Gaps of one or more residues can beinserted into a query sequence, a subject sequence, or both, to maximizesequence alignments. For fast pairwise alignment of nucleic acidsequences, the following default parameters are used: word size: 2;window size: 4; scoring method: percentage; number of top diagonals: 4;and gap penalty: 5. For multiple alignment of nucleic acid sequences,the following parameters are used: gap opening penalty: 10.0; gapextension penalty: 5.0; and weight transitions: yes. For fast pairwisealignment of protein sequences, the following parameters are used: wordsize: 1; window size: 5; scoring method: percentage; number of topdiagonals: 5; and gap penalty: 3. For multiple alignment of proteinsequences, the following parameters are used: weight matrix: blosum; gapopening penalty: 10.0; gap extension penalty: 0.05; hydrophilic gaps:on; hydrophilic residues: Gly, Pro, Ser, Asn, Asp, Gln, Glu, Arg, andLys; and residue-specific gap penalties: on. The output is a sequencealignment that reflects the relationship between sequences. ClustalW canbe run, for example, at the Baylor College of Medicine Search Launchersite (searchlauncher.bcm.tmc.edu/multi-align/multi-align.html) and atthe European Bioinformatics Institute site on the World Wide Web(ebi.ac.uk/clustalw). To determine a “percent identity” between a querysequence and a subject sequence, the number of matching bases or aminoacids in the alignment is divided by the total number of matched andmismatched bases or amino acids, followed by multiplying the result by100.

It is noted that the percent identity value can be rounded to thenearest tenth. For example, 78.11, 78.12, 78.13, and 78.14 are roundeddown to 78.1, while 78.15, 78.16, 78.17, 78.18, and 78.19 are rounded upto 78.2. It also is noted that the length value will always be aninteger.

Recombinant Constructs, Vectors and Host Cells

Vectors containing nucleic acids such as those described herein also areprovided. A “vector” is a replicon, such as a plasmid, phage, or cosmid,into which another DNA segment may be inserted so as to bring about thereplication of the inserted segment. Generally, a vector is capable ofreplication when associated with the proper regulatory regions. Suitablevector backbones include, for example, those routinely used in the artsuch as plasmids, viruses, artificial chromosomes, BACs, YACs, or PACs.The term “vector” includes cloning and expression vectors, as well asviral vectors and integrating vectors. An “expression vector” is avector that includes one or more regulatory regions. Suitable expressionvectors include, without limitation, plasmids and viral vectors derivedfrom, for example, bacteriophage, baculoviruses, tobacco mosaic virus,herpesviruses, cytomegalovirus, vaccinia viruses, adenoviruses,adeno-associated viruses, and retroviruses. Numerous vectors andexpression systems are commercially available from such corporations asNovagen (Madison, Wis.), Clontech (Palo Alto, Calif.), Stratagene (LaJolla, Calif.), and Invitrogen/Life Technologies (Carlsbad, Calif.).

The term “regulatory region” refers to nucleotide sequences thatinfluence transcription or translation initiation and rate, andstability and/or mobility of the transcript or polypeptide product.Regulatory regions include, without limitation, promoter sequences,enhancer sequences, response elements, protein recognition sites,inducible elements, promoter control elements, protein bindingsequences, 5′ and 3′ untranslated regions (UTRs), transcriptional startsites, termination sequences, polyadenylation sequences, introns, andother regulatory regions that can reside within coding sequences, suchas secretory signals and protease cleavage sites.

As used herein, the term “operably linked” refers to positioning of aregulatory region and a transcribable sequence in a nucleic acid so asto allow or facilitate transcription of the transcribable sequence. Forexample, a regulatory region is operably linked to a coding sequencewhen RNA polymerase is able to transcribe the coding sequence into mRNA,which then can be translated into a protein encoded by the codingsequence.

Promoters are involved in recognition and binding of RNA polymerase andother proteins to initiate and modulate transcription. To bring a codingsequence under the control of a promoter, it typically is necessary toposition the translation initiation site of the translational readingframe of the polypeptide between one and about fifty nucleotidesdownstream of the promoter. A promoter can, however, be positioned asmuch as about 5,000 nucleotides upstream of the translation start site,or about 2,000 nucleotides upstream of the transcription start site. Apromoter typically comprises at least a core (basal) promoter. Apromoter also may include at least one control element such as anupstream element. Such elements include upstream activation regions(UARs) and, optionally, other DNA sequences that affect transcription ofa polynucleotide such as a synthetic upstream element. The choice ofpromoters to be included depends upon several factors, including, butnot limited to, efficiency, selectability, inducibility, desiredexpression level, and cell or tissue specificity.

Constitutive Promoters

Constitutive promoters can promote transcription of an operably linkednucleic acid under most, but not necessarily all, environmentalconditions and states of development or cell differentiation.Non-limiting examples of constitutive promoters that can be included inthe nucleic acid constructs provided herein include the cauliflowermosaic virus (CaMV) 35S transcription initiation region, the mannopinesynthase (MAS) promoter, the 1′ or 2′ promoters derived from T-DNA ofAgrobacterium tumefaciens, the figwort mosaic virus 35S promoter, actinpromoters such as the rice actin promoter, ubiquitin promoters such asthe maize ubiquitin-1 promoter, p32449, and p13879.

Broadly Expressing Promoters

A promoter can be said to be “broadly expressing” when it promotestranscription in many, but not all, plant tissues. For example, abroadly expressing promoter can promote transcription of an operablylinked sequence in one or more of the stem, shoot, shoot tip (apex), andleaves, but can promote transcription weakly or not at all in tissuessuch as reproductive tissues of flowers and developing seeds. In certaincases, a broadly expressing promoter operably linked to a sequence canpromote transcription of the linked sequence in a plant shoot at a levelthat is at least two times, e.g., at least 3, 5, 10, or 20 times,greater than the level of transcription in a developing seed. In othercases, a broadly expressing promoter can promote transcription in aplant shoot at a level that is at least two times, e.g., at least 3, 5,10, or 20 times, greater than the level of transcription in areproductive tissue of a flower. In view of the above, the CaMV 35Spromoter is not considered a broadly expressing promoter. Non-limitingexamples of broadly expressing promoters that can be included in thenucleic acid constructs provided herein include the p326, YP0158,YP0214, YP0380, PT0848, PTO633, YP0050, YP0144 and YP0190 promoters.See, e.g., U.S. patent application Ser. No. 11/208,308, filed Aug. 19,2005.

Tissue-, organ- and cell-specific promoters confer transcription only orpredominantly in a particular tissue, organ, and cell type,respectively. In some embodiments, promoters specific to non-seedtissues, such as vegetative tissues, can be suitable regulatory regions.Vegetative tissues include the stem, parenchyma, ground meristem,vascular bundle, cambium, phloem, cortex, shoot apical meristem, lateralshoot meristem, root apical meristem, lateral root meristem, leafprimordium, leaf mesophyll, or leaf epidermis.

Root-Specific Promoters

Root-specific promoters confer transcription only or predominantly inroot tissue. Examples of root-specific promoters include the rootspecific subdomains of the CaMV 35S promoter (Lam et al., Proc Natl AcadSci USA 86:7890-7894 (1989)), root cell specific promoters reported byConkling et al. Plant Physiol. 93:1203-1211 (1990), and the tobacco RD2gene promoter.

Seed-Specific Promoters

In some embodiments, promoters that are essentially specific to seedscan be useful. Transcription from a seed-specific promoter occursprimarily in endosperm and cotyledon tissue during seed development.Non-limiting examples of seed-specific promoters that can be included inthe nucleic acid constructs provided herein include the napin promoter,the Arcelin-5 promoter, the phaseolin gene promoter (Bustos et al.,Plant Cell 1(9):839-853 (1989)), the soybean trypsin inhibitor promoter(Riggs et al., Plant Cell 1(6):609-621 (1989)), the ACP promoter(Baerson et al., Plant Mol Biol, 22(2):255-267 (1993)), the stearoyl-ACPdesaturase gene (Slocombe et al., Plant Physiol 104(4): 167-176 (1994)),the soybean α′ subunit of β-conglycinin promoter (Chen et al., Proc.Natl. Acad Sci. U.S.A. 83:8560-8564 (1986)), the oleosin promoter (Honget al., Plant Mol Biol 34(3):549-555 (1997)), zein promoters such as the15 kD zein promoter, the 16 kD zein promoter, 19 kD zein promoter, 22 kDzein promoter and 27 kD zein promoter. Also suitable are the Osgt-1promoter from the rice glutelin-1 gene (Zheng et al., Mol. Cell Biol.13:5829-5842 (1993)), the beta-amylase gene promoter, and the barleyhordein gene promoter.

Non-Seed Fruit Tissue Promoters

Promoters that are active in non-seed fruit tissues can also be useful,e.g., a polygalacturonidase promoter, the banana TRX promoter, and themelon actin promoter.

Female Gametophyte Specific Promoters

To achieve female gametophyte specific expression, regulatory elementsthat preferentially drive transcription in female gametophytic tissuesare used, such as embryo sac promoters. Most suitable are regulatoryelements that preferentially drive transcription in polar nuclei or thecentral cell, or in precursors to polar nuclei, but not in egg cells orprecursors to egg cells. A regulatory element whose pattern oftranscription extends from polar nuclei into early endosperm developmentis also acceptable, although rapidly diminishing transcription inendosperm tissue after fertilization is most preferred. Expression inthe zygote or developing embryo is not preferred.

Female reproductive tissue promoters that may be suitable include thosederived from the following genes: maize MAC1 (see, Sheridan (1996)Genetics, 142:1009-1020); maize Cat3 (see, GenBank No. L05934; Abler(1993) Plant Mol. Biol., 22:10131-1038); Arabidopsis viviparous-1 (see,Genbank No. U93215); Arabidopsis atmycl (see, Urao (1996) Plant Mol.Biol., 32:571-57; Conceicao (1994) Plant, 5:493-505).

Other female gametophyte tissue promoters include those derived from thefollowing genes: Arabidopsis Fie (GenBank No. AF129516); ArabidopsisMea; and Arabidopsis Fis2 (GenBank No. AF096096); ovule BEL1 (Reiser(1995) Cell, 83:735-742; Ray (1994) Proc. Natl. Acad Sci. U.S.A.,91:5761-5765; GenBank No. U39944); Fie 1.1 (U.S. Pat. No. 6,906,244) andArabidopsis DMC1 (see, GenBank No. U76670). Ovary-specific promotersinclude the tomato pz7 gene promoter and the tomato pz130 gene promoter.Other exemplary female gametophyte tissue-specific promoters include thefollowing Arabidopsis promoters: YP0039, YP0101, YP0102, YP0110, YP0117,YP0119, YP0137, DME PROMOTER, YP0285 and YP0212. Female gametophytetissue promoters that may be useful in monocotyledonous plants such asrice include the following promoters: Y678g10, p756a09, Y790g04,p780a10, Y730e07, Y760g09, p530c10, p524d05, p523d11 and p472e10.

Photosynthetically-Active Tissue Promoters

Photosynthetically-active tissue promoters confer transcription only orpredominantly in photosynthetically active tissue. Examples of suchpromoters include the ribulose-1,5-bisphosphate carboxylase (RbcS)promoters such as the RbcS promoter from eastern larch (Larix laricina),the pine cab6 promoter (Yamamoto et al., Plant Cell Physiol. 35:773-778(1994)), the Cab-1 gene promoter from wheat (Fejes et al., Plant Mol.Biol. 15:921-932 (1990)), the CAB-1 promoter from spinach (Lubberstedtet al., Plant Physiol. 104:997-1006 (1994)), the cab1R promoter fromrice (Luan et al., Plant Cell 4:971-981 (1992)), the pyruvate,orthophosphate dikinase (PPDK) promoter from corn (Matsuoka et al.,Proc. Natl. Acad. Sci. U.S.A. 90:9586-9590 (1993)), the tobacco Lhcb1*2promoter (Cerdan et al., Plant Mol. Biol. 33:245-255 (1997)), theArabidopsis thaliana SUC2 sucrose-H+ symporter promoter (Truernit etal., Planta. 196:564-570 (1995)), and thylakoid membrane proteinpromoters from spinach (psaD, psaF, psaE, PC, FNR, atpC, atpD, cab,rbcS).

Basal Promoters

A basal promoter is the minimal sequence necessary for assembly of atranscription complex required for transcription initiation. Basalpromoters frequently include a “TATA box” element that may be locatedbetween about 15 and about 35 nucleotides upstream from the site oftranscription initiation. Basal promoters also may include a “CCAAT box”element (typically the sequence CCAAT) and/or a GGGCG sequence, whichcan be located between about 40 and about 200 nucleotides, typicallyabout 60 to about 120 nucleotides, upstream from the transcription startsite.

Other Promoters

Other classes of promoters include, but are not limited to, induciblepromoters, such as promoters that confer transcription in response toexternal stimuli such as chemical agents, developmental stimuli, orenvironmental stimuli.

Other suitable promoters include those set forth in U.S. PatentApplication Ser. Nos. 60/505,689; 60/518,075; 60/544,771; 60/558,869;60/583,691; 60/619,181; 60/637,140; 10/957,569; 11/058,689; 11/172,703and PCT/US05/23639, e.g., promoters designated YP0086 (gDNA ID 7418340),YP0188 (gDNA ID 7418570), YP0263 (gDNA ID 7418658), p13879, p32449,PT0758; PT0743; PT0829; YP0096 and YP0119.

Other Regulatory Regions

A 5′ untranslated region (UTR) is transcribed, but is not translated,and lies between the start site of the transcript and the translationinitiation codon and may include the +1 nucleotide. A 3′ UTR can bepositioned between the translation termination codon and the end of thetranscript. UTRs can have particular functions such as increasing mRNAmessage stability or translation attenuation. Examples of 3′ UTRsinclude, but are not limited to polyadenylation signals andtranscription termination sequences.

A polyadenylation region at the 3′-end of a coding region can also beoperably linked to a coding sequence. The polyadenylation region can bederived from the natural gene, from various other plant genes, or fromtransfer-DNA (T-DNA).

A suitable enhancer is a cis-regulatory element (−212 to −154) from theupstream region of the octopine synthase (ocs) gene. Fromm et al., ThePlant Cell 1:977-984 (1989).

The vectors provided herein also can include, for example, origins ofreplication, scaffold attachment regions (SARs), and/or markers. Amarker gene can confer a selectable phenotype on a plant cell. Forexample, a marker can confer, biocide resistance, such as resistance toan antibiotic (e.g., kanamycin, G418, bleomycin, or hygromycin), or anherbicide (e.g., chlorosulfuron or phosphinothricin). In addition, anexpression vector can include a tag sequence designed to facilitatemanipulation or detection (e.g., purification or localization) of theexpressed polypeptide. Tag sequences, such as green fluorescent protein(GFP), glutathione S-transferase (GST), polyhistidine, c-myc,hemagglutinin, or Flag™ tag (Kodak, New Haven, Conn.) sequencestypically are expressed as a fusion with the encoded polypeptide. Suchtags can be inserted anywhere within the polypeptide, including ateither the carboxyl or amino terminus.

It will be understood that more than one regulatory region may bepresent in a recombinant polynucleotide, e.g., introns, enhancers,upstream activation regions, and inducible elements. Thus, more than oneregulatory region can be operably linked to the sequence encoding asugar-modulating polypeptide.

The recombinant DNA constructs provided herein typically include apolynucleotide sequence (e.g., a sequence encoding a sugar-modulatingpolypeptide) inserted into a vector suitable for transformation of plantcells. Recombinant vectors can be made using, for example, standardrecombinant DNA techniques (see, e.g., Sambrook et al. (1989) MolecularCloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory,Cold Spring Harbor, N.Y.).

Transgenic Plants and Cells

Any of the vectors provided herein can be used to transform plant cellsand, if desired, generate transgenic plants. Thus, transgenic plants andplant cells containing the nucleic acids described herein also areprovided, as are methods for making such transgenic plants and plantcells. A plant or plant cells can be transformed by having the constructintegrated into its genome, i.e., can be stably transformed. Stablytransformed cells typically retain the introduced nucleic acid sequencewith each cell division. Alternatively, the plant or plant cells alsocan be transiently transformed such that the construct is not integratedinto its genome. Transiently transformed cells typically lose some orall of the introduced nucleic acid construct with each cell division,such that the introduced nucleic acid cannot be detected in daughtercells after sufficient number of cell divisions. Both transientlytransformed and stably transformed transgenic plants and plant cells canbe useful in the methods described herein.

Typically, transgenic plant cells used in the methods described hereinconstitute part or all of a whole plant. Such plants can be grown in amanner suitable for the species under consideration, either in a growthchamber, a greenhouse, or in a field. Transgenic plants can be bred asdesired for a particular purpose, e.g., to introduce a recombinantnucleic acid into other lines, to transfer a recombinant nucleic acid toother species, or for further selection of other desirable traits.Alternatively, transgenic plants can be propagated vegetatively forthose species amenable to such techniques. Progeny includes descendantsof a particular plant or plant line. Progeny of an instant plant includeseeds formed on F₁, F₂, F₃, F₄, F₅, F₆, and subsequent generationplants, or seeds formed on BC₁, BC₂, BC₃, and subsequent generationplants, or seeds formed on F₁BC₁, F₁BC₂, F₁BC₃, and subsequentgeneration plants. Seeds produced by a transgenic plant can be grown andthen selfed (or outcrossed and selfed) to obtain seeds homozygous forthe nucleic acid construct.

Alternatively, transgenic plant cells can be grown in suspensionculture, or tissue or organ culture, for production of secondarymetabolites. For the purposes of the methods provided herein, solidand/or liquid tissue culture techniques can be used. When using solidmedium, transgenic plant cells can be placed directly onto the medium orcan be placed onto a filter film that is then placed in contact with themedium. When using liquid medium, transgenic plant cells can be placedonto a floatation device, e.g., a porous membrane that contacts theliquid medium. Solid medium typically is made from liquid medium byadding agar. For example, a solid medium can be Murashige and Skoog (MS)medium containing agar and a suitable concentration of an auxin, e.g.,2,4-dichlorophenoxyacetic acid (2,4-D), and a suitable concentration ofa cytokinin, e.g., kinetin.

Techniques for transforming a wide variety of higher plant species areknown in the art. The polynucleotides and/or recombinant vectorsdescribed herein can be introduced into the genome of a plant host usingany of a number of known methods, including electroporation,microinjection, and biolistic methods. Alternatively, polynucleotides orvectors can be combined with suitable T-DNA flanking regions andintroduced into a conventional Agrobacterium tumefaciens host vector.Such Agrobacterium tumefaciens-mediated transformation techniques,including disarming and use of binary vectors, are well known in theart. Other gene transfer and transformation techniques includeprotoplast transformation through calcium or PEG,electroporation-mediated uptake of naked DNA, electroporation of planttissues, viral vector-mediated transformation, and microprojectilebombardment (see, e.g., U.S. Pat. Nos. 5,538,880; 5,204,253; 5,591,616;and 6,329,571). If a cell or tissue culture is used as the recipienttissue for transformation, plants can be regenerated from transformedcultures using techniques known to those skilled in the art.

The polynucleotides and vectors described herein can be used totransform a number of monocotyledonous and dicotyledonous plants andplant cell systems, including dicots such as alfalfa, amaranth, apple,beans (including kidney beans, lima beans, dry beans, green beans),broccoli, cabbage, carrot, castor bean, chick peas, cherry, chicory,chocolate, clover, coffee, cotton, cottonseed, crambe, eucalyptus, flax,grape, grapefruit, lemon, lentils, lettuce, linseed, mango, melon (e.g.,watermelon, cantaloupe), mustard, orange, peanut, peach, pear, peas,pepper, plum, poplar, potato, rapeseed (high erucic acid and canola),safflower, sesame, soybean, spinach, strawberry, sugarbeet, sunflower,tea, tomato, as well as monocots such as banana, barley, date palm,field corn, garlic, millet, oat, oil palm, onion, pineapple, popcorn,rice, rye, sorghum, sudangrass, sugarcane, sweet corn, switchgrass, turfgrasses, and wheat. Gymnosperms such as fir, pine and spruce can also besuitable. Brown seaweeds, green seaweeds, red seaweeds, and microalgaealso can be used.

Thus, the methods and compositions described herein can be used withdicotyledonous plants belonging, for example, to the ordersAristochiales, Asterales, Batales, Campanulales, Capparales,Caryophyllales, Casuarinales, Celastrales, Cornales, Diapensales,Dilleniales, Dipsacales, Ebenales, Ericales, Eucomiales, Euphorbiales,Fabales, Fagales, Gentianales, Geraniales, Haloragales, Hamamelidales,Illiciales, Juglandales, Lamiales, Laurales, Lecythidales, Leitneriales,Magniolales, Malvales, Myricales, Myrtales, Nymphaeales, Papeverales,Piperales, Plantaginales, Plumbaginales, Podostemales, Polemoniales,Polygalales, Polygonales, Primulales, Proteales, Rafflesiales,Ranunculales, Rhamnales, Rosales, Rubiales, Salicales, Santales,Sapindales, Sarraceniaceae, Scrophulariales, Trochodendrales, Theales,Umbellales, Urticales, and Violales. The methods and compositionsdescribed herein also can be utilized with monocotyledonous plants suchas those belonging to the orders Alismatales, Arales, Arecales,Bromeliales, Commelinales, Cyclanthales, Cyperales, Eriocaulales,Hydrocharitales, Juncales, Lilliales, Najadales, Orchidales, Pandanales,Poales, Restionales, Triuridales, Typhales, Zingiberales, and withplants belonging to Gymnospermae, e.g., Pinales, Ginkgoales, Cycadalesand Gnetales.

The methods and compositions can be used over a broad range of plantspecies, including species from the dicot genera Apium, Alseodaphne,Anacardium, Arabidopsis, Arachis, Atropa, Beilschmiedia, Bixa, Brassica,Capsicum, Calendula, Carthamus, Chicorium, Cinnamomum, Citrus,Citrullus, Cocculus, Cocos, Coffea, Corylus, Croton, Cucumis, Cucurbita,Cuphea, Daucus, Dianthus, Duguetia, Euphoria, Ficus, Fragaria, Glaucium,Glycine, Glycyrrhiza, Gossypium, Helianthus, Hevea, Hyoscyamus, Lactuca,Landolphia, Linum, Litsea, Lycopersicon, Lupinus, Majorana, Malus,Manihot, Medicago, Mentha, Nicotiana, Olea, Parthenium, Persea, Petunia,Phaseolus, Pistacia, Pisum, Populus sect., Prunus, Pyrus, Raphanus,Ricinus, Rosa, Rosmarinus, Rubus, Salix, Senecio, Sinapis, Solanum,Spinacia, Stephania, Tagetes, Theobroma, Trifolium, Trigonella,Vaccinium, Vicia, Vigna, Vitis; and the monocot genera Alliuin,Andropogon, Ananus, Aragrostis, Asparagus, Avena, Cynodon, Elaeis,Festuca, Festulolium, Heterocallis, Hordeum, Lemna, Lolium, Musa, Oryza,Panicum, Pannesetum, Phleum, Poa, Phoenix, Saccharum, Secale, Sorghum,Triticum, and Zea; and the gymnosperm genera Abies, Cunninghamia, Picea,and Pinus.

The methods and compositions described herein also can be used withbrown seaweeds, e.g., Ascophyllum nodosum, Fucus vesiculosus, Fucusserratus, Himanthalia elongata, and Undaria pinnatifida; red seaweeds,e.g., Porphyra umbilicalis, Palmaria palmata, Cracilaria verrucosa, andChondrus crispus; green seaweeds, e.g., Ulva spp. and Enteromorpha spp.;and microalgae, e.g., Spirulina sp. (S. platensis and S. maxima) andOdontella aurita. In addition, the methods and compositions can be usedwith Crypthecodinium cohnii, Schizochytrium spp., and Haematococcuspluvialis.

In some embodiments, a plant can be from a species selected from Ananuscomosus, Arabidopsis thaliana, Brassica rapa, Brassica napus, Brassicaoleracea, Bixa orellana, Calendula officinalis, Cinnamomum camphora,Coffea arabica, Glycine max, Glycyrrhiza glabra, Gossypium hirsutum,Gossypium herbaceum, Lactuca sativa, Lycopersicon esculentum, Menthapiperita, Mentha spicata, Musa paradisiaca, Oryza sativa, Partheniumargentatum, Rosmarinus officinalis, Solanum tuberosum, Theobroma cacao,Triticum aestivum, Vitis vinifera, and Zea mays. For example, in certainembodiments, plants from the following species can be preferred: Ananuscomosus, Brassica rapa, Brassica napus, Brassica oleracea, Coffeaarabica, Glycine max, Gossypium hirsutum, Gossypium herbaceum, Lactucasativa, Lycopersicon esculentum, Mentha piperita, Mentha spicata, Musaparadisiaca, Oryza Sativa, Parthenium argentatum, Solanum tuberosum,Theobroma cacao, Triticum aestivum, Vitis vinifera, and Zea mays.

A transformed cell, callus, tissue, or plant can be identified andisolated by selecting or screening the engineered plant material forparticular traits or activities, e.g., those encoded by marker genes orantibiotic resistance genes. Such screening and selection methodologiesare well known to those having ordinary skill in the art. In addition,physical and biochemical methods can be used to identify transformants.These include Southern analysis or PCR amplification for detection of apolynucleotide; Northern blots, S1 RNase protection, primer-extension,quantitative real-time PCR, or reverse transcriptase PCR (RT-PCR)amplification for detecting RNA transcripts; enzymatic assays fordetecting enzyme or ribozyme activity of polypeptides andpolynucleotides; and protein gel electrophoresis, Western blots,immunoprecipitation, and enzyme-linked immunoassays to detectpolypeptides. Other techniques such as in situ hybridization, enzymestaining, and immunostaining also can be used to detect the presence orexpression of polypeptides and/or polynucleotides. Methods forperforming all of the referenced techniques are well known. After apolynucleotide is stably incorporated into a transgenic plant, it can beintroduced into other plants using, for example, standard breedingtechniques.

Transgenic plants (or plant cells) can have an altered phenotype ascompared to a corresponding control plant (or plant cell) that eitherlacks the transgene or does not express the transgene. A polypeptide canaffect the phenotype of a plant (e.g., a transgenic plant) whenexpressed in the plant, e.g., at the appropriate time(s), in theappropriate tissue(s), or at the appropriate expression levels.Phenotypic effects can be evaluated relative to a control plant thatdoes not express the exogenous polynucleotide of interest, such as acorresponding wild-type plant, a corresponding plant that is nottransgenic for the exogenous polynucleotide of interest but otherwise isof the same genetic background as the transgenic plant of interest, or acorresponding plant of the same genetic background in which expressionof the polypeptide is suppressed, inhibited, or not induced (e.g., whereexpression is under the control of an inducible promoter). A plant canbe said “not to express” a polypeptide when the plant exhibits less than10 percent (e.g., less than 9 percent, 8 percent, 7 percent, 6 percent,5 percent, 4 percent, 3 percent, 2 percent, 1 percent, 0.5 percent, 0.1percent, 0.01 percent, or 0.001 percent) of the amount of polypeptide ormRNA encoding the polypeptide exhibited by the plant of interest.Expression can be evaluated using methods including, for example,quantitative real-time PCR, RT-PCR, Northern blots, SI RNase protection,primer extensions, Western blots, protein gel electrophoresis,immunoprecipitation, enzyme-linked immunoassays, chip assays, and massspectrometry. It should be noted that if a polypeptide is expressedunder the control of a tissue-specific or broadly expressing promoter,expression can be evaluated in the entire plant or in a selected tissue.Similarly, if a polypeptide is expressed at a particular time, e.g., ata particular time in development or upon induction, expression can beevaluated selectively at a desired time period.

When a sugar-modulating polypeptide described herein is expressed in aplant, the transgenic plant can have an increased level of one or moresugars (e.g., glucose, fructose, or sucrose). For example, non-seedtissues of a transgenic plant can exhibit increased levels of one ormore of glucose, fructose, and/or sucrose. The sugar level can beincreased by at least 5 percent (e.g., 5, 10, 15, 20, 25, 30, 35, 40,45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120,125, 130, 135, 140, 145, 150, 175, 200, 225, 250, 275, 300, 350, 400,450, 500, 550, 600, 650, 700, 800, 900, or 1000 percent) as compared tothe sugar level in a corresponding control plant that does not expressthe transgene. For example, a level of glucose, fructose, or sucrose innon-seed tissues of a plant can be increased by at least 7 percent toabout 120 percent or any value therebetween, such as at least 9 percent,10 percent, 11 percent, 15 percent, 18 percent, 20 percent, 21 percent,22 percent, 24 percent, 27 percent, 29 percent, 30 percent, 31 percent,35 percent, 36 percent, 37 percent, 40 percent, 43 percent, 50 percent,51 percent, 60 percent, 63 percent, 70 percent, 75 percent, 80 percent,90 percent, 105 percent, or 115 percent, as compared to thecorresponding levels in a control plant. In some cases, a level ofglucose in non-seed tissues of a plant can be increased by at least 10percent to about 120 percent or any value therebetween, such as at least15 percent, 18 percent, 21 percent, 24 percent, 27 percent, 29 percent,31 percent, 35 percent, 37 percent, 40 percent, 43 percent, 50 percent,60 percent, 70 percent, 75 percent, 80 percent, 90 percent, 105 percent,or 115 percent, as compared to the corresponding levels in a controlplant. In some cases, a level of fructose in non-seed tissues of a plantcan be increased by at least 7 percent to about 115 percent or any valuetherebetween, such as at least 10 percent, 15 percent, 18 percent, 20percent, 24 percent, 30 percent, 36 percent, 40 percent, 44 percent, 50percent, 55 percent, 60 percent, 70 percent, 75 percent, 80 percent, 90percent, 105 percent, or 110 percent, as compared to the correspondinglevels in a control plant. In other cases, a level of sucrose innon-seed tissues of a plant can be increased by at least 10 percent toabout 100 percent or any value therebetween, such as at least 12percent, 15 percent, 18 percent, 24 percent, 29 percent, 35 percent, 40percent, 45 percent, 51 percent, 56 percent, 60 percent, 65 percent, 70percent, 75 percent, 80 percent, 85 percent, 92 percent, or 97 percent,as compared to the corresponding levels in a control plant. In yet othercases, levels of glucose and fructose in non-seed tissues of a plant canbe increased by at least 7 percent to about 120 percent or any valuetherebetween, such as at least 10 percent, 12 percent, 15 percent, 18percent, 20 percent, 21 percent, 24 percent, 27 percent, 29 percent, 30percent, 31 percent, 35 percent, 36 percent, 37 percent, 40 percent, 43percent, 50 percent, 60 percent, 70 percent, 75 percent, 80 percent, 90percent, 105 percent, or 115 percent, as compared to the correspondinglevels in a control plant.

Seeds, Extracts, Non-Seed Tissues, Animal Feed, and Articles ofManufacture

Also provided herein are compositions such as food and feed products,and articles of manufacture, such as bags of seeds, based on transgenicplants described herein. Typically, a substantially uniform mixture ofseeds is conditioned and bagged in packaging material by means known inthe art to form an article of manufacture. Packaging material such aspaper and cloth are well known in the art. Such a bag of seed preferablyhas a package label accompanying the bag, e.g., a tag or label securedto the packaging material, a label printed on the packaging material, ora label inserted within the bag. The package label may indicate thatseed contained therein incorporates transgenes that provide increasedamounts of one or more sugars in one or more tissues of plants grownfrom such seeds.

Seeds from transgenic plants described herein can be used as is, e.g.,to grow plants, or can be used to make food products such as flours,vegetable oils, and insoluble fibers. Non-seed tissues from transgenicplants described herein can be used as is or can be used to make foodproducts such as fresh, canned, and frozen fruits and vegetables. Seedsand non-seed tissues from transgenic plants described herein also can beused as animal feed. Transgenic plants described herein also can be usedto make grains, such as wheat, oat, rice, barley, quinoa, and rye. Suchproducts are useful to provide increased amounts of sugar(s) in the dietand to provide increased flavor.

Transgenic plants described herein can also serve as raw materialssuitable for fermentation to produce ethanol. For example, corn cobs,corn stalks, sugarcane, sugarbeets, fruit (fresh or dried), citrusmolasses, cane sorghum, and cotton can be fermented to produce ethylalcohol. Fuel ethanol can also be manufactured using sugarcane juice ormolasses as raw material. Producing ethanol from plant materialscontaining increased amounts of sugar can improve ethanol yields.

Transgenic plants described herein can also be used as a source fromwhich to extract sugars, using techniques known in the art. For example,sugar can be extracted from sugarcane, sugarbeet, date palm, sorghum,and sugar maple. Molasses can also be extracted from sugarcane. Theresulting extracts can be purified. Purified sugar, sugar syrup, sugarjuice, or extracts containing sugar can be included in nutritionalsupplements as well as processed food products, e.g., soft drinks,sports drinks, ice cream, baked goods, relishes, sauces, tomato paste,canned foods, meats, salads, candy, fruit juices, vegetable juices,syrup, snack products, frozen entrees, breakfast cereals, breakfastbars, baby foods, and high fructose corn syrup. Sugar can also beincluded in cell culture media.

Methods

Also provided herein are methods that employ the describedpolynucleotides, polypeptides, plant cells, transgenic plants, seeds,and tissues. For example, a method of modulating the level of a sugar ina plant, such as non-seed tissue or seeds of a plant, is provided. Themethod includes introducing one or more exogenous nucleic acidsdescribed herein into a plant cell. A modulated level can be anincreased level of a sugar, including one or more of glucose, fructose,and/or sucrose.

A method of producing a plant having a modulated sugar level (e.g., anincreased glucose, fructose, and/or sucrose level) is also provided,which includes introducing into a plant cell one or more exogenousnucleic acids as previously described, and growing a plant from theplant cell. The increased level of one or more of glucose, fructose,and/or sucrose can be in the seed and/or the non-seed tissue of theplant.

A method of producing a sugar is also provided. The method includesextracting sugar from a transgenic plant (e.g., sugarcane or sugarbeet)described herein. Sugar can be extracted from such plants usingtechniques known in the art.

Transgenic plants (e.g., corn, wheat, and sugarbeets) having increasedsugar levels can also be useful in lactate/lactic acid productionprocesses. Lactate can be used to produce polylactide polymers (see, forexample, U.S. Pat. No. 6,291,597). Furthermore, sugars can be useful inthe production of polylactide polymers.

Finally, a method of producing ethanol is provided. The method includesfermentation of plant materials based on transgenic plants providedherein. Plant materials can be fermented to produce ethanol usingtechniques known in the art (see, for example, U.S. Pat. Nos. 6,509,180and 6,927,048).

When the polynucleotides and polypeptides provided herein are expressednon-naturally (e.g., with respect to location in a plant, such as rootvs. stem; environmental condition; plant species; time of development;and/or expression level), they can produce plants with modulated levelsof sugars. These traits can be used to make use of or maximize plantproducts, including, without limitation, non-seed plant tissues, roots,seeds, flowers, fruits, extracts, and oils. For example, nucleic acidsprovided herein can be used to generate transgenic plants havingincreased expression of one or more polynucleotides involved in sugarsynthesis. In some cases, nucleic acids provided herein can be used togenerate transgenic plants having increased or decreased expression ofone or more polypeptides involved in maintenance of sugar levels (e.g.,regulation of sugar degradation). In some cases, nucleic acids providedherein can be used to generate transgenic plants having increasedexpression of one or more polypeptides involved in regulating expressionof one or more genes involved in sugar synthesis or maintenance of sugarlevels. Such transgenic plants may produce higher levels of one or moresugars (e.g., glucose, fructose, or sucrose), as discussed herein. Thus,the polynucleotides and polypeptides provided herein can be useful inthe preparation of transgenic plants having particular application inthe agricultural and nutritional industries.

The invention will be further described in the following examples, whichdo not limit the scope of the invention described in the claims.

EXAMPLES Example 1 Transgenic Plants

The following symbols are used in the Examples: T₁: first generationtransformant; T₂: second generation, progeny of self-pollinated T₁plants; T₃: third generation, progeny of self-pollinated T₂ plants; T₄:fourth generation, progeny of self-pollinated T₃ plants. Independenttransformations are referred to as events.

Ceres clone 625627 (SEQ ID NO:13) contains a nucleotide sequence thathas homology to two different nucleotide sequences. The first 510nucleotides at the 5′ end correspond to a portion of an expressedsequence tag (GenBank accession number AW734757) from soybean thatcontains an AP2 DNA-binding domain. Nucleotides 505-725 correspond to aportion of a nucleic acid sequence that encodes an ATPase relatedprotein. Ceres clone 625627 encodes a 174 amino acid polypeptide.

Ceres clone 32380 (SEQ ID NO:1) encodes a 232 amino acid polypeptidepredicted to be a DNA-directed RNA polymerase II third largest subunit.

Ti plasmid vectors were constructed that contained Ceres clone 625627 orCeres clone 32380 operably linked to the 35S promoter. The Ti plasmidvector used for these constructs, CRS338, contained a phosphinothricinacetyltransferase gene, which confers Finale™ resistance to transformedplants. Wild-type Arabidopsis Wassilewskija (Ws) plants were transformedseparately with each Ti plasmid vector, essentially as described inBechtold et al., C.R. Acad. Sci. Paris, 316:1194-1199 (1993).

Arabidopsis lines containing Ceres clone 625627 or Ceres clone 32380were designated ME02225 or ME05896, respectively. The presence of theCeres clone 625627 vector in ME02225, and the Ceres clone 32380 vectorin ME05896, was confirmed by Finale™ resistance, PCR amplification fromgreen leaf tissue extract, and sequencing of PCR products. As controls,wild-type Arabidopsis Wassilewskija (Ws) plants were transformed withthe empty vector SR00559.

Ten events of each of ME02225 and ME05896 were selected and screened forvisible phenotypic alterations in the T₁ generation. The physicalappearance of all T₁ plants was identical to that of the control plants.

Example 2 Analysis of Sugar Levels in ME02225 Events

Plants were grown from a mixture of seeds collected from T₁ events ofME02225. The plants were harvested ten days post-bolting. Non-seedtissues (e.g., aerial tissues) from four segregating Finale™-resistantT₂ plants were pooled and immediately frozen in liquid nitrogen. Thetissues were stored at −80° C. and subsequently lyophilized for 72hours. The freeze-dried tissues were crushed into a fine powder andprepared for analysis using gas chromatography-mass spectroscopy(GC-MS). Briefly, the freeze-dried tissues were extracted in triplicateusing methanol, and then extracted using dichloromethane. The polarphases were derivatized using transmethylation, methoxyamination, andtrimethylsylation. Derivatized extracts (2 μL) were injected into aShimadzu GC-MS QP-2010 (Shimadzu Scientific Instruments, Columbia, Md.).The data were analyzed using the Shimadzu GC-MS Solutions software(Shimadzu Scientific Instruments). Briefly, target ion peak areas wereintegrated after identity confirmation using retention time standardsand reference ion peak ratios. The target ion peak areas were normalizedwith respect to the internal standard and compared relative to thecontrol sample. The normalized peak areas from glucose and fructose wereaveraged and the standard deviations were calculated.

Non-seed tissues from T₂ plants of ME02225 had increased levels ofglucose and fructose compared to the levels of glucose and fructose innon-seed tissues from corresponding control plants. As presented in FIG.5, the levels of glucose and fructose were increased by 105% and 90%,respectively, in T₂ plants of ME02225 compared to the levels of glucoseand fructose in the corresponding control plants.

In addition to analyzing ME02225 plants grown from a mixture of seeds,events of ME02225 were analyzed individually. Seeds from each of fourevents of ME02225 were planted separately. T₂ and T₃ plants from each ofthe four events of ME02225 were grown until ten days post-bolting.Non-seed tissues from four Finale™-resistant plants of each event werepooled, frozen in liquid nitrogen, and stored at −.-80° C. The frozentissues were lyophilized for 72 hours and stored at −80° C. Thefreeze-dried tissues were crushed into a fine powder and prepared foranalysis using GC-MS. Briefly, the lyophilized plant tissues wereextracted in triplicate using methanol, and then extracted usingdichloromethane. The polar phases were derivatized using methoxyamineand N-Methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA). Derivatizedextracts were analyzed using GC-MS, and the data were analyzed asdescribed above.

The GC-MS analysis showed that Finale™-resistant T₂ plants from events-02 and -04 had significantly increased glucose levels compared tocontrol plants. As presented in Table 1, glucose levels were increasedby 24% and 35% in events -02 and -04, respectively, compared to thecorresponding control plants. TABLE 1 Glucose levels (% Control) in T₂and T₃ plants from ME02225 events Ws ME02225- ME02225- ME02225- ME02225-Control 02 03 04 05 T₂ 100 ± 12 124 ± 3 109 ± 21 135 ± 8   77 ± 3 p- NA0.07 0.56 0.02 0.08 value T₃ 100 ± 12 137 ± 6 108 ± 7  129 ± 11 103 ± 4p- NA <0.01  0.28 0.02 0.61 value

Levels of glucose in Finale™-resistant T₃ plants from four ME02225events also were analyzed using GC-MS. Events -02 and -04 hadsignificantly increased glucose levels compared to control plants. Aspresented in Table 1, glucose levels were increased by 37% and 29% inevents -02 and -04, respectively, compared to the corresponding controlplants.

The GC-MS analysis also showed that Finale™-resistant T₂ plants fromevents -02 and -04 had significantly increased fructose levels comparedto control plants. As presented in Table 2, fructose levels wereincreased by 10% and 30% in events -02 and -04, respectively, comparedto the corresponding control plants. The 10% increase in fructose levelin event -02 was not statistically significant. TABLE 2 Fructose levels(% Control) in T₂ and T₃ plants from ME02225 events Ws ME02225- ME02225-ME02225- ME02225- Control 02 03 04 05 T₂ 100 ± 17 110 ± 8  101 ± 15 130± 6   70 ± 6 p- NA 0.41 0.92 0.08 0.08 value T₃ 100 ± 13 140 ± 10 110 ±12 130 ± 10 100 ± 9 p- NA <0.01  0.45 <0.01  <0.01  value

Levels of fructose in Finale™-resistant T₃ plants from four ME02225events also were analyzed using GC-MS. Events -02 and -04 hadsignificantly increased fructose levels compared to control plants. Aspresented in Table 2, fructose levels were increased by 40% and 30% inevents -02 and -04, respectively, compared to the corresponding controlplants.

There were no observable or statistically significant differencesbetween T₂ ME02225 and control plants in germination, onset offlowering, rosette diameter, fertility, plant height, and generalmorphology/architecture.

Example 3 Analysis of Sugar Levels in ME05896 Events

Levels of sucrose in Finale™-resistant T₂ plants from five ME05896events were analyzed using GC-MS, as described above. As presented inTable 3, sucrose levels were increased by 63% and 22% in events -01 and-02, respectively, compared to the corresponding control plants.

Levels of sucrose in Finale™-resistant T₃ plants from five ME05896events also were analyzed using GC-MS. As presented in Table 3, thetrend of increased sugar level exhibited by T₃ plants of ME05896 issimilar to that exhibited by the T₂ plants, particularly in the case ofevent -01. TABLE 3 Sucrose levels (fold increase) in T₂ plants fromME05896 events ME05896-01 ME05896-02 ME05896-03 ME05896-04 ME05896-10Control T₂ 1.63 ± 0.12 1.22 ± 0.07 1.07 ± 0.04 1.01 ± 0.08 1.07 ± 0.081.00 ± 0.13 p-value <0.01  0.08 0.40 0.90 0.48 NA T₃ 1.14 ± 0.09 1.01 ±0.03 1.07 ± 0.03 1.02 ± 0.05 0.89 ± 0.03 1.00 ± 0.06 p-value 0.10 0.400.01 0.40 0.01 NA

Example 4 Determination of Functional Homolog and/or Ortholo Sequences

A subject sequence was considered a functional homolog and/or orthologof a query sequence if the subject sequence encoded a protein having afunction and/or activity similar to the protein encoded by the querysequence. A process known as Reciprocal BLAST (Rivera et al., Proc.Natl. Acad. Sci. (U.S.A.), 1998, 95:6239-6244) was used to identifypotential functional homolog and/or ortholog sequences from availabledatabases of public and proprietary peptide sequences, including theNCBI NR protein database and a private Ceres database of peptidetranslations of sequences from Ceres clones.

Before starting a Reciprocal BLAST process, BLAST was used to search aspecific query polypeptide against all polypeptides from its sourcespecies in order to identify polypeptides having 80% or greater sequenceidentity with the query polypeptide. The query polypeptide together withpolypeptides identified as having 80% or greater sequence identity withthe query polypeptide were designated as a cluster.

The main Reciprocal BLAST process consists of two rounds of BLASTsearches: a forward search and a reverse search. In the forward searchstep, a query polypeptide sequence, “polypeptide A,” from source speciesSA was BLASTed against all protein sequences from a species of interest.The best matches, or top hits, were determined using an E-value cutoffof 10⁻⁵ and an identity cutoff of 35%. Among the top hits, the hit withthe lowest E-value was considered the best hit and a potentialfunctional homolog and/or ortholog. Any other top hit(s) having 80% orgreater sequence identity with the best hit or the original querypolypeptide was also considered a potential functional homolog and/orortholog. This process was repeated for all species of interest.

In the reverse search of the Reciprocal Blast process, the top hitsidentified in the forward search, from all species, were BLASTed againstall protein sequences from the source species SA. A top hit from theforward search that returned a polypeptide from the aforementionedcluster as its best hit was also considered a potential functionalhomolog and/or ortholog.

Manual inspection of potential functional homologs and/or orthologs wascarried out to select identified functional homologs and/or orthologs.The results are presented in FIG. 6. Percent identities to SEQ ID NO:2are shown in Table 4 below. SEQ ID % Iden- Designation Species NO: tityCeresClone 32380 Arabidopsis thaliana 2 100.00 gi|21593370 Arabidopsisthaliana 3 86.80 CeresClone 692249 Glycine max 4 76.40 CeresClone 244359Zea mays 6 73.50 gi|50898416 Oryza sativa subsp. japonica 7 72.40CeresClone 698259 Triticum aestivum 5 70.10 gi|21593370T Arabidopsisthaliana 8 87.00 CeresClone692249T Glycine max 9 76.02 CeresClone244359TZea mays 11 70.00 gi|50898416T Oryza sativa subsp. japonica 12 68.97CeresClone 698259T Triticum aestivum 10 66.52

OTHER EMBODIMENTS

It is to be understood that while the invention has been described inconjunction with the detailed description thereof, the foregoingdescription is intended to illustrate and not limit the scope of theinvention, which is defined by the scope of the appended claims. Otheraspects, advantages, and modifications are within the scope of thefollowing claims.

1. A method of modulating the level of a sugar in a plant, said methodcomprising introducing into a plant cell an isolated nucleic acidcomprising a nucleic acid sequence encoding a polypeptide having 80percent or greater sequence identity to an amino acid sequence selectedfrom the group consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, Ceres clone SEQ IDNO:9, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, and theConsensus sequence set forth in FIG. 6, wherein a plant produced fromsaid plant cell has a different sugar level as compared to a sugar levelin a corresponding control plant that does not comprise said isolatednucleic acid.
 2. The method of claim 1, wherein said isolated nucleicacid comprises a nucleic acid sequence encoding a polypeptide having 85percent or greater sequence identity to an amino acid sequence selectedfrom the group consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ IDNO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, and the Consensus sequenceset forth in FIG.
 6. 3. The method of claim 1, wherein said isolatednucleic acid comprises a nucleic acid sequence encoding a polypeptidehaving 90 percent or greater sequence identity to an amino acid sequenceselected from the group consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ IDNO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9,SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, and the Consensussequence set forth in FIG.
 6. 4. The method of claim 1, wherein saidisolated nucleic acid comprises a nucleic acid sequence encoding apolypeptide having 95 percent or greater sequence identity to an aminoacid sequence selected from the group consisting of SEQ ID NO:14, SEQ IDNO:2, SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4,SEQ ID NO:9, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, andthe Consensus sequence set forth in FIG.
 6. 5. A method of modulatingthe level of a sugar in a plant, said method comprising introducing intoa plant cell an isolated nucleic acid comprising a nucleic acid sequenceencoding a polypeptide having 80 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ IDNO:11, SEQ ID NO:4, SEQ ID NO:9, and the Consensus sequence set forth inFIG. 6, wherein a plant produced from said plant cell has a differentsugar level as compared to a sugar level in a corresponding controlplant that does not comprise said isolated nucleic acid.
 6. The methodof claim 5, wherein said isolated nucleic acid comprises a nucleic acidsequence encoding a polypeptide having 85 percent or greater sequenceidentity to an amino acid sequence selected from the group consisting ofSEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQID NO:11, SEQ ID NO:4, SEQ ID NO:9, and the Consensus sequence set forthin FIG.
 6. 7. The method of claim 5, wherein said isolated nucleic acidcomprises a nucleic acid sequence encoding a polypeptide having 90percent or greater sequence identity to an amino acid sequence selectedfrom the group consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, and theConsensus sequence set forth in FIG.
 6. 8. The method of claim 5,wherein said isolated nucleic acid comprises a nucleic acid sequenceencoding a polypeptide having 95 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ IDNO:11, SEQ ID NO:4, SEQ ID NO:9, and the Consensus sequence set forth inFIG.
 6. 9. A method of modulating the level of a sugar in a plant, saidmethod comprising introducing into a plant cell an isolated nucleic acidcomprising a nucleic acid sequence encoding a polypeptide having 80percent or greater sequence identity to an amino acid sequence selectedfrom the group consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQID NO:6, SEQ ID NO:4, and the Consensus sequence set forth in FIG. 6,wherein a plant produced from said plant cell has a different sugarlevel as compared to a sugar level in a corresponding control plant thatdoes not comprise said isolated nucleic acid.
 10. The method of claim 9,wherein said isolated nucleic acid comprises a nucleic acid sequenceencoding a polypeptide having 85 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:4, and theConsensus sequence set forth in FIG.
 6. 11. The method of claim 9,wherein said isolated nucleic acid comprises a nucleic acid sequenceencoding a polypeptide having 90 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:4, and theConsensus sequence set forth in FIG.
 6. 12. The method of claim 9,wherein said isolated nucleic acid comprises a nucleic acid sequenceencoding a polypeptide having 95 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:4, and theConsensus sequence set forth in FIG.
 6. 13. A method of modulating thelevel of a sugar in a plant, said method comprising introducing into aplant cell an isolated nucleic acid comprising a nucleic acid sequenceencoding a polypeptide having 80 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:4, wherein aplant produced from said plant cell has a different sugar level ascompared to a sugar level in a corresponding control plant that does notcomprise said isolated nucleic acid.
 14. The method of claim 13, whereinsaid isolated nucleic acid comprises a nucleic acid sequence encoding apolypeptide having 85 percent or greater sequence identity to an aminoacid sequence selected from the group consisting of SEQ ID NO:14, SEQ IDNO:2, SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:4.
 15. The method of claim13, wherein said isolated nucleic acid comprises a nucleic acid sequenceencoding a polypeptide having 90 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:4.
 16. Themethod of claim 13, wherein said isolated nucleic acid comprises anucleic acid sequence encoding a polypeptide having 95 percent orgreater sequence identity to an amino acid sequence selected from thegroup consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6,and SEQ ID NO:4.
 17. A method of modulating the level of a sugar in aplant, said method comprising introducing into a plant cell an isolatednucleic acid comprising a nucleic acid sequence encoding a polypeptidecomprising an amino acid sequence corresponding to SEQ ID NO:2, whereina plant produced from said plant cell has a different sugar level ascompared to a sugar level in a corresponding control plant that does notcomprise said isolated nucleic acid.
 18. A method of modulating thelevel of a sugar in a plant, said method comprising introducing into aplant cell an isolated nucleic acid comprising a nucleic acid sequenceencoding a polypeptide comprising an amino acid sequence correspondingto SEQ ID NO:14, wherein a plant produced from said plant cell has adifferent sugar level as compared to a sugar level in a correspondingcontrol plant that does not comprise said isolated nucleic acid.
 19. Amethod of modulating the level of a sugar in a plant, said methodcomprising introducing into a plant cell an isolated nucleic acidcomprising a nucleic acid sequence encoding a polypeptide comprising anamino acid sequence corresponding to the Consensus sequence set forth inFIG. 6, wherein a plant produced from said plant cell has a differentsugar level as compared to a sugar level in a corresponding controlplant that does not comprise said isolated nucleic acid.
 20. A method ofmodulating the level of a sugar in a plant, said method comprisingintroducing into a plant cell (a) a first isolated nucleic acidcomprising a nucleic acid sequence encoding a polypeptide having 80percent or greater sequence identity to an amino acid sequence selectedfrom the group consisting of SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ IDNO:12, SEQ ID NO:3, SEQ ID NO:8, and the Consensus sequence set forth inFIG. 6; and (b) a second isolated nucleic acid comprising a nucleic acidsequence encoding a polypeptide having 80 percent or greater sequenceidentity to an amino acid sequence corresponding to SEQ ID NO:14;wherein a plant produced from said plant cell has a different sugarlevel as compared to a sugar level in a corresponding control plant thatdoes not comprise said first isolated nucleic acid or said secondisolated nucleic acid.
 21. A method of modulating the level of a sugarin a plant, said method comprising introducing into a plant cell (a) afirst isolated nucleic acid comprising a nucleic acid sequence encodinga polypeptide comprising an amino acid sequence selected from the groupconsisting of SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQID NO:4, SEQ ID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ IDNO:3, SEQ ID NO:8, and the Consensus sequence set forth in FIG. 6; and(b) a second isolated nucleic acid comprising a nucleic acid sequenceencoding a polypeptide comprising an amino acid sequence correspondingto SEQ ID NO:14; wherein a plant produced from said plant cell has adifferent sugar level as compared to a sugar level in a correspondingcontrol plant that does not comprise said first isolated nucleic acid orsaid second isolated nucleic acid.
 22. A method of modulating the levelof a sugar in a plant, said method comprising introducing into a plantcell (a) a first isolated nucleic acid comprising a nucleic acidsequence encoding a polypeptide having 80 percent or greater sequenceidentity to an amino acid sequence selected from the group consisting ofSEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, and SEQ IDNO:8; and (b) a second isolated nucleic acid comprising a nucleic acidsequence encoding a polypeptide having 80 percent or greater sequenceidentity to an amino acid sequence corresponding to SEQ ID NO:14;wherein a plant produced from said plant cell has a different sugarlevel as compared to a sugar level in a corresponding control plant thatdoes not comprise said first isolated nucleic acid or said secondisolated nucleic acid.
 23. A method of modulating the level of a sugarin a plant, said method comprising introducing into a plant cell (a) afirst isolated nucleic acid comprising a nucleic acid sequence encodinga polypeptide comprising an amino acid sequence selected from the groupconsisting of SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQID NO:4, SEQ ID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ IDNO:3, and SEQ ID NO:8; and (b) a second isolated nucleic acid comprisinga nucleic acid sequence encoding a polypeptide comprising an amino acidsequence corresponding to SEQ ID NO:14; wherein a plant produced fromsaid plant cell has a different sugar level as compared to a sugar levelin a corresponding control plant that does not comprise said firstisolated nucleic acid or said second isolated nucleic acid.
 24. Themethod of claim 1, 5, 9, 13, 17, 18, 19, 20, 21, 22, or 23, wherein saiddifferent sugar level is an increased level of one or more sugars. 25.The method of claim 24, wherein said sugar is glucose.
 26. The method ofclaim 24, wherein said sugar is fructose.
 27. The method of claim 24,wherein said sugar is sucrose.
 28. The method of claim 24, wherein saiddifferent sugar level is an increased level of glucose and fructose. 29.The method of claim 24, wherein said different sugar level is anincreased level of glucose, fructose, and sucrose.
 30. A method ofproducing a plant having a modulated level of a sugar, said methodcomprising (a) introducing into a plant cell an isolated nucleic acidcomprising a nucleic acid sequence encoding a polypeptide having 80percent or greater sequence identity to an amino acid sequence selectedfrom the group consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ IDNO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, and the Consensus sequenceset forth in FIG. 6; and (b) growing a plant from said plant cell. 31.The method of claim 30, wherein said isolated nucleic acid comprises anucleic acid sequence encoding a polypeptide having 85 percent orgreater sequence identity to an amino acid sequence selected from thegroup consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ IDNO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:7,SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, and the Consensus sequence setforth in FIG.
 6. 32. The method of claim 30, wherein said isolatednucleic acid comprises a nucleic acid sequence encoding a polypeptidehaving 90 percent or greater sequence identity to an amino acid sequenceselected from the group consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ IDNO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9,SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, and the Consensussequence set forth in FIG.
 6. 33. The method of claim 30, wherein saidisolated nucleic acid comprises a nucleic acid sequence encoding apolypeptide having 95 percent or greater sequence identity to an aminoacid sequence selected from the group consisting of SEQ ID NO:14, SEQ IDNO:2, SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4,SEQ ID NO:9, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, andthe Consensus sequence set forth in FIG.
 6. 34. A method of producing aplant having a modulated level of a sugar, said method comprising (a)introducing into a plant cell an isolated nucleic acid comprising anucleic acid sequence encoding a polypeptide having 80 percent orgreater sequence identity to an amino acid sequence selected from thegroup consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ IDNO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, and theConsensus sequence set forth in FIG. 6; and (b) growing a plant fromsaid plant cell.
 35. The method of claim 34, wherein said isolatednucleic acid comprises a nucleic acid sequence encoding a polypeptidehaving 85 percent or greater sequence identity to an amino acid sequenceselected from the group consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ IDNO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9,and the Consensus sequence set forth in FIG.
 6. 36. The method of claim34, wherein said isolated nucleic acid comprises a nucleic acid sequenceencoding a polypeptide having 90 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ IDNO:11, SEQ ID NO:4, SEQ ID NO:9, and the Consensus sequence set forth inFIG.
 6. 37. The method of claim 34, wherein said isolated nucleic acidcomprises a nucleic acid sequence encoding a polypeptide having 95percent or greater sequence identity to an amino acid sequence selectedfrom the group consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, and theConsensus sequence set forth in FIG.
 6. 38. A method of producing aplant having a modulated level of a sugar, said method comprising (a)introducing into a plant cell an isolated nucleic acid comprising anucleic acid sequence encoding a polypeptide having 80 percent orgreater sequence identity to an amino acid sequence selected from thegroup consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6,SEQ ID NO:4, and the Consensus sequence set forth in FIG. 6; and (b)growing a plant from said plant cell.
 39. The method of claim 38,wherein said isolated nucleic acid comprises a nucleic acid sequenceencoding a polypeptide having 85 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:4, and theConsensus sequence set forth in FIG.
 6. 40. The method of claim 38,wherein said isolated nucleic acid comprises a nucleic acid sequenceencoding a polypeptide having 90 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:4, and theConsensus sequence set forth in FIG.
 6. 41. The method of claim 38,wherein said isolated nucleic acid comprises a nucleic acid sequenceencoding a polypeptide having 95 percent or greater sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:4, and theConsensus sequence set forth in FIG.
 6. 42. A method of producing aplant having a modulated level of a sugar, said method comprising (a)introducing into a plant cell an isolated nucleic acid comprising anucleic acid sequence encoding a polypeptide having 80 percent orgreater sequence identity to an amino acid sequence selected from thegroup consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6,and SEQ ID NO:4; and (b) growing a plant from said plant cell.
 43. Themethod of claim 42, wherein said isolated nucleic acid comprises anucleic acid sequence encoding a polypeptide having 85 percent orgreater sequence identity to an amino acid sequence selected from thegroup consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:6,and SEQ ID NO:4.
 44. The method of claim 42, wherein said isolatednucleic acid comprises a nucleic acid sequence encoding a polypeptidehaving 90 percent or greater sequence identity to an amino acid sequenceselected from the group consisting of SEQ ID NO:14, SEQ ID NO:2, SEQ IDNO:5, SEQ ID NO:6, and SEQ ID NO:4.
 45. The method of claim 42, whereinsaid isolated nucleic acid comprises a nucleic acid sequence encoding apolypeptide having 95 percent or greater sequence identity to an aminoacid sequence selected from the group consisting of SEQ ID NO:14, SEQ IDNO:2, SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:4.
 46. A method ofproducing a plant having a modulated level of a sugar, said methodcomprising (a) introducing into a plant cell an isolated nucleic acidcomprising a nucleic acid sequence encoding a polypeptide comprising anamino acid sequence corresponding to SEQ ID NO:14; and (b) growing aplant from said plant cell.
 47. A method of producing a plant having amodulated level of a sugar, said method comprising (a) introducing intoa plant cell an isolated nucleic acid comprising a nucleic acid sequenceencoding a polypeptide comprising an amino acid sequence correspondingto SEQ ID NO:2; and (b) growing a plant from said plant cell.
 48. Amethod of producing a plant having a modulated level of a sugar, saidmethod comprising (a) introducing into a plant cell an isolated nucleicacid comprising a nucleic acid sequence encoding a polypeptidecomprising an amino acid sequence corresponding to the Consensussequence set forth in FIG. 6; and (b) growing a plant from said plantcell.
 49. A method of producing a plant having a modulated level of asugar, said method comprising (a) introducing into a plant cell a firstisolated nucleic acid comprising a nucleic acid sequence encoding apolypeptide having 80 percent or greater sequence identity to an aminoacid sequence selected from the group consisting of SEQ ID NO:5, SEQ IDNO:10, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:2,SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:3, SEQ ID NO:8, and the Consensussequence set forth in FIG. 6, and a second isolated nucleic acidcomprising a nucleic acid sequence encoding a polypeptide having 80percent or greater sequence identity to an amino acid sequencecorresponding to SEQ ID NO:14; and (b) growing a plant from said plantcell.
 50. A method of producing a plant having a modulated level of asugar, said method comprising (a) introducing into a plant cell a firstisolated nucleic acid comprising a nucleic acid sequence encoding apolypeptide comprising an amino acid sequence selected from the groupconsisting of SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQID NO:4, SEQ ID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ IDNO:3, SEQ ID NO:8, and the Consensus sequence set forth in FIG. 6, and asecond isolated nucleic acid comprising a nucleic acid sequence encodinga polypeptide comprising an amino acid sequence corresponding to SEQ IDNO:14; and (b) growing a plant from said plant cell.
 51. A method ofproducing a plant having a modulated level of a sugar, said methodcomprising (a) introducing into a plant cell a first isolated nucleicacid comprising a nucleic acid sequence encoding a polypeptide having 80percent or greater sequence identity to an amino acid sequence selectedfrom the group consisting of SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQID NO:11, SEQ ID NO:4, SEQ ID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ IDNO:12, SEQ ID NO:3, and SEQ ID NO:8, and a second isolated nucleic acidcomprising a nucleic acid sequence encoding a polypeptide having 80percent or greater sequence identity to an amino acid sequencecorresponding to SEQ ID NO:14; and (b) growing a plant from said plantcell.
 52. A method of producing a plant having a modulated level of asugar, said method comprising (a) introducing into a plant cell a firstisolated nucleic acid comprising a nucleic acid sequence encoding apolypeptide comprising an amino acid sequence selected from the groupconsisting of SEQ ID NO:5, SEQ ID NO:10, SEQ ID NO:6, SEQ ID NO:11, SEQID NO:4, SEQ ID NO:9, SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ IDNO:3, and SEQ ID NO:8, and a second isolated nucleic acid comprising anucleic acid sequence encoding a polypeptide comprising an amino acidsequence corresponding to SEQ ID NO:14; and (b) growing a plant fromsaid plant cell.
 53. The method of claim 30, 34, 38, 42, 46, 47, 48, 49,50, 51, or 52, wherein said modulated level is an increased level of oneor more sugars.
 54. The method of claim 53, wherein said sugar isglucose.
 55. The method of claim 53, wherein said sugar is fructose. 56.The method of claim 53, wherein said sugar is sucrose.
 57. The method ofclaim 53, wherein said modulated level is an increased level of glucoseand fructose.
 58. The method of claim 53, wherein said modulated levelis an increased level of glucose, fructose, and sucrose.
 59. The methodof claim 1, 5, 9, 13, 17, 18, 19, 30, 34, 38, 42, 46, 47, or 48, whereinsaid isolated nucleic acid is operably linked to a regulatory region.60. The method of claim 20, 21, 22, 23, 49, 50, 51, or 52, wherein saidfirst isolated nucleic acid and said second isolated nucleic acid isoperably linked to a regulatory region.
 61. The method of claim 59 or60, wherein said regulatory region is a promoter.
 62. The method ofclaim 61, wherein said promoter is a cell-specific or tissue-specificpromoter.
 63. The method of claim 61, wherein said promoter is a broadlyexpressing promoter.
 64. The method of claim 63, wherein said broadlyexpressing promoter is selected from the group consisting of p326,YP0158, YP0214, YP0380, PT0848, PT0633, YP0050, YP0144, and YP0190. 65.The method of claim 62, wherein said tissue-specific promoter is aseed-specific promoter.
 66. The method of claim 65, wherein saidseed-specific promoter is selected from the group consisting of thenapin promoter, the Arcelin-5 promoter, the phaseolin gene promoter, thesoybean trypsin inhibitor promoter, the ACP promoter, the stearoyl-ACPdesaturase gene, the soybean α′ subunit of β-conglycinin promoter, theoleosin promoter, the 15 kD zein promoter, the 16 kD zein promoter, the19 kD zein promoter, the 22 kD zein promoter, the 27 kD zein promoter,the Osgt-1 promoter, the beta-amylase gene promoter, and the barleyhordein gene promoter.
 67. The method of claim 62, wherein saidtissue-specific promoter is a root-specific promoter.
 68. The method ofclaim 67, wherein said root-specific promoter is selected from the groupconsisting of the root specific subdomains of the CaMV 35S promoter andthe tobacco RD2 gene promoter.
 69. The method of claim 62, wherein saidtissue-specific promoter is a non-seed fruit tissue promoter.
 70. Themethod of claim 69, wherein said non-seed fruit tissue promoter isselected from the group consisting of a polygalacturonidase promoter,the banana TRX promoter, and the melon actin promoter.
 71. The method ofclaim 61, wherein said promoter is a constitutive promoter.
 72. Themethod of claim 71, wherein said promoter is selected from the groupconsisting of 35S, p32449, and p13879.
 73. The method of claim 61,wherein said promoter is an inducible promoter.
 74. The method of claim1, 5, 9, 13, 17, 18, 19, 20, 21, 22, 23, 30, 34, 38, 42, 46, 47, 48, 49,50, 51, or 52, wherein said plant is from a genus selected from thegroup consisting of Abies, Agrostis, Allium, Alseodaphne, Anacardium,Ananus, Andropogon, Arachis, Apium, Arabidopsis, Aragrostis,Ascophyllum, Asparagus, Atropa, Avena, Beilschmiedia, Bixa, Brassica,Calendula, Capsicum, Carthamus, Chondrus, Chicorium, Cinnamomum, Citrus,Citrullus, Cocculus, Cocos, Coffea, Corylus, Cracilaria, Croton,Crypthecodinium, Cucumis, Cucurbita, Cunninghamia, Cuphea, Cynodon,Daucus, Dianthus, Duguetia, Elaeis, Enteromorpha, Euphoria, Festuca,Festulolium, Ficus, Fragaria, Fucus, Glaucium, Glycine, Glycyrrhiza,Gossypium, Haematococcus, Helianthus, Heterocallis, Hevea, Himanthalia,Hordeum, Hyoscyamus, Lactuca, Landolphia, Lemna, Linum, Litsea, Lolium,Lycopersicon, Lupinus, Majorana, Malus, Manihot, Medicago, Mentha, Musa,Nicotiana, Odontella, Olea, Oryza, Palmaria, Panicum, Pannesetum,Parthenium, Persea, Petunia, Phaseolus, Phleum, Phoenix, Picea, Pinus,Pistacia, Pisum, Poa, Populus sect., Porphyra, Prunus, Pyrus, Raphanus,Ricinus, Rosa, Rosmarinus, Rubus, Saccharum, Salix, Schizochytrium,Secale, Senecio, Sinapis, Solanum, Sorghum, Spinacia, Spirulina,Stephania, Triticum, Tagetes, Theobroma, Trifolium, Trigonella, Ulva,Undaria, Vaccinium, Vicia, Vigna, Vitis, and Zea.
 75. The method ofclaim 1, 5, 9, 13, 17, 18, 19, 20, 21, 22, 23, 30, 34, 38, 42, 46, 47,48, 49, 50, 51, or 52, wherein said plant is a species selected fromAnanus comosus, Arabidopsis thaliana, Brassica rapa, Brassica napus,Brassica oleracea, Bixa orellana, Calendula officinalis, Cinnamomumcamphora, Coffea arabica, Glycine max, Glycyrrhiza glabra, Gossypiumhirsutum, Gossypium herbaceum, Lactuca sativa, Lycopersicon esculentum,Mentha piperita, Mentha spicata, Musa paradisiaca, Oryza sativa,Parthenium argentatum, Rosmarinus officinalis, Solanum tuberosum,Theobroma cacao, Triticum aestivum, Vitis vinifera, and Zea mays. 76.The method of claim 1, 5, 9, 13, 17, 18, 19, 20, 21, 22, 23, 30, 34, 38,42, 46, 47, 48, 49, 50, 51, or 52, wherein said plant is selected fromthe group consisting of alfalfa, amaranth, apple, beans (includingkidney beans, lima beans, dry beans, green beans), broccoli, cabbage,carrot, castor bean, chick peas, cherry, chicory, chocolate, clover,coffee, cotton, cottonseed, crambe, eucalyptus, flax, grape, grapefruit,lemon, lentils, lettuce, linseed, mango, melon (e.g., watermelon,cantaloupe), mustard, orange, peanut, peach, pear, peas, pepper, plum,poplar, potato, rapeseed (high erucic acid and canola), safflower,sesame, soybean, spinach, strawberry, sugarbeet, sunflower, tea, tomato,banana, barley, date palm, field corn, garlic, millet, oat, oil palm,onion, pineapple, popcorn, rice, rye, sorghum, sudangrass, sugarcane,sweet corn, switchgrass, turf grasses, wheat, fir, pine, spruce, brownseaweeds, green seaweeds, red seaweeds, and microalgae.